saqarTvelos agraruli universiteti - NPLG...adamianis sisxlis Srats unari aqvs moaxdinos maltis...

saqarTvelos agraruli universiteti

eliso mamisaSvili

M

msxvilfexa pirutyvSi brucelozis serodiagnostika da

paTogenis genetikuri analizi

veterinariis doqtoris akademiuri

xarisxis mosapoveblad warmodgenili

d i s e r t a c i a

specialoba: saveterinaro mikrobiologia, virusologia, epizootologia,

mikologia, imunologia, parazitologia

xelmZRvaneli :

veterinariis mecnierebaTa doqtori,

sruli profesori merab naTiZe

T b i l i s i

2 0 1 2

2

s a r C e v i

1. S e s a v a l i --------------------------------------------------------------------------- 4

2. literaturis mimoxilva

2.1. brucelozi. istoria, gavrceleba sasoflo-sameurneo

cxovelebSi.--------------------------------------------------------------------------------------- 12

2.2. brucelebis nairsaxeobebi, migracia cxovelebSi,

paTogenezi. -------------------------------------------------------------------------------------- 17

2.3. veterinariasa da medicinaSi brucelozis serologi-

uri da genetikuri diagnostika. --------------------------------------------------- 25

3. sakuTari gamokvlevebi

3.1. gamokvlevis masala da meTodebi. ------------------------------------------ 38

3.2. saqarTveloSi 2005-2011w.w. msxvilfexa pirutyvis

bruceloziT daavadebis dinamika.----------------------------------------- 44

3.3. msxvilfexa pirutyvis sisxlis Sratebis roz-bengal sinjiT

da konkurentuli imunofermentuli analiziT gamokvlevis

Sedegebi. ---------------------------------------------------------------------------------- 55

3.4. furebis rZis brucelozze rgoluri reaqciiT gamokvlevis

Sedegebi. -------------------------------------------------------------------------------- 63

3.5. furebidan gamoyofili B. abortus Seswavlis Sedegebi.---------- 68

3.5.1. morfologia da tinqtorialuri Tvisebebi. ----------------------- 70

3

3.5.2. sakveb areebze zrdis Tavisebureba. ----------------------------------- 72

3.5.3. B. abortus izolatebis bioqimiuri Tvisebebi. ----------------------- 77

3.5.4. B. abortus bovis izolatebis fagoidentifikacia. ------------------ 85

3.5.5. B. abortus bovis izolatebis d.n.m skriningi. --------------------------- 89

4. miRebuli Sedegebis analizi. ------------------------------------------- 98

5. d a s k v n e b i. ------------------------------------------------------------- 105

6. praqtikuli winadadebebi. -------------------------------------------------- 107

7. gamoyenebuli literatura. ------------------------------------------------- 108

4

1. S e s a v a l i

Temis aqtualoba: `brucelozi momavlis daavadebaa”…es

gamonaTqvami ekuTvnis frang mecniers Sarl nikols, romelic

meoce saukunis dasawyisSi warmoTqva parizSi. brucelozi 180

nozoologiur zoonozur infeqciebs Soris, dRemde rCeba

aqtualur problemad. (В. М. Авилов с соавт, 1997; А. Д. Антоненко,

2000;. M. L. Boschiroli, 2001,). brucelozi cxovelebSi da adamianebSi

jer kidev yovelmxriv ar aris Seswavlili.

janmrTelobis dacvisa da saveterinaro medicinis

TvalsazrisiT brucelozi warmoadgens msoflio problemas. (M.

J. Corbel, 1997, M. L. Boschiroli, 2001).Ees daavadeba mecxoveleobas

udides zarals ayenebs. Bbrucelozi cxovelebidan gadadis

adamianze dainficirebuli sakvebi produqtebiT, inhalaciuri an

daavadebul cxovelTan pirdapiri kontaqtiT da iwveves mZime,

xangrZlivad mimdinare daavadebas. (R. Hurtado. 2001.), rac

brucelozTan brZolas did socialur mniSvnelobas aniWebs.

amasTan mxedvelobaSia misaRebi ekonomikuri mxarec. daavadebuli

adamiani didi xniT kargavs Sromis unars, sWirdeba mkurnaloba.

adamianebi, romlebic uSualod uvlian daavadebul pirutyvs

(mwvelavebi, mwyemsebi da a.S.) saWiroebs profilaqtikur

saSualebaTa gamoyenebas. (j. babikiSvili, m. mamaiaSvili. Tanaavt.

2005).

cxovelTa dacvis msoflio organizaciis (OIE) monacemebiT,

dedamiwis xuTive kontinentis 193 saxelmwifodan mxolod 38-ia

Tavisufali brucelozisagan; xuTSi infeqcia dafiqsirebulia

gamonaklisi SemTxvevebis saxiT. ormocdaaT qveyanaSi daavadebis

erTeuli SemTxvevebia aRwerili. enzootiuri gavrcelebis for-

5

miT brucelozi gvxvdeba 38 saxelmwifoSi, xolo eqvsSi

poulobs farTe gavrcelebas; gansakuTrebiT xmelTaSua zRvis

auzSi, mcire aziaSi, arabeTis yureSi, meqsikaSi, centralur da

samxreT amerikaSi (Z. Getikua et. al., 2005; A. Alim, Z. D. Tomul, 2005;)

aRmosavleT aziaSi, afrikaSi.

samxreT amerikis qveynebSi, kanadaSi, a.S.S-s TiTqmis yvela

StatSi gvxvdeba brucelozi da weliwadSi 100-200 SemTxvevaa

fiqsirebuli. aRsaniSnavia, rom a.S.S-s CrdiloeTis StatebSi

ZiriTadad registrirebulia msxvilfexa pirutyvis brucelozi.

(T. J. Doyle, R. T Bryan., 2000.). samxreT amerikis qveynebidan

msxvilfexa pirutyvSi brucelozi registrirebulia braziliaSi,

argentinaSi, urugvaiSi, CileSi (Lopez Merino A, et. al. 1989., Dr. Alfredo

Garín, et. al. 2011.,) msxvilfexa pirutyvis brucelozi

registrirebulia avstraliaSi, axal zelandiaSi (W. L McDonald,

et. al. 2006.).

saudis arabeTSi romelic warmoadgens maRal endemur zonas,

yovelwliurad fiqsirebulia adamianis bruceloziT daavadebis

8000 SemTxveva (Z. Memish, 2001).

zogierTma qveynebma, gansakuTrebiT evropulma, (inglisi, dania,

germania, fineTi, Svecia, norvegia, Sveicaria, CexeTi, slovenia,

rumineTi), agreTve iaponiam miaRwia brucelozis srul

likvidacias cxovelebSi da adamianebSi. (S Al Dahouk, et. al. 2002., K. E.

Bergsrom et. al. 2003., S. Krkic-Dautovic et. al. 2006). brucelozTan

warmatebulad mimdinareobs brZola bulgareTSi, yofili

iugoslaviis qveynebSi, sadac daavadebis erTeuli SemTxvevebia

registrirebuli. (R. D. Jones et. al., 2004; Z. Cvetnic,

et. al., 2003).

brucelozze yuradReba gansakuTrebiT gamaxvildaBbolo

6

periodSi, kerZod brucelebi gamoyofil iqna zRvis

ZuZumwovrebidan, (sur. 1) romlebic aavadeben xmeleTis

ZuZumwovrebs da adamiansac, rac qmnis SesaZlo ekologiur

problemas. (S. D. Bre.; L. L. Perrett; et. al., 1999; A. R. Spickler, J. A. Roth, et. al.,

2010;)

sur. 1. zRvis ZuZumwovrebi.

7

brucelebi, gansakuTrebiT saSiSi gaxda, a.S.S-Si 2001 wlis 11

seqtembris tragikuli movlenebisa da mis Semdgom ganviTarebuli

bioteraqtebis seriebis Semdeg, rogorc SesaZlo biologiuri

iaraRi. daavadebaTa kontrolis centris (CDC), mier brucelebi

miekuTvneba ,,b” kategorias (Centers for Disease Control and Prevantion.

2000B.). ,,b” kategoriis agentebi SedarebiT swrafad vrceldeba.

brucelozis aRmZvreli arabeTisa da zogierTi qveynebis

arsenalSia, rogorc biologiuri iaraRi. (D. Shocham 2000, R. A.

Greenfield, et. al, 2002., A. R. Spickler, J. A. Roth, et. al., 2010.).

brucelozi enzootiuri daavadebaa, rac gulisxmobs mis

gavrcelebas mxolod gansazRvrul adgilebSi, meurneobebSi,

fermebSi, dasaxlebul punqtebSi. is SeiZleba gaCndes wlis

nebismier dros.Mmas axasiaTebs infeqciuri procesis stadiuroba

(latenturi-usimptomo, regionaluri, generalizebuli) nayofisa

da Sardsasqeso sistemebis organoebis inficireba, ujredSida da

organosSida parazitizmi, rac aisaxeba imunitetis arasteriluri-

infeqciuri da steriluri-arainfeqciuri fazebis gamovlinebaSi.

brucelozTan brZolis sirTule ganpirobebulia

daavadebis aRmZvrelis mravali saxeobis arsebobiT da maTi

SesaZlo migraciiT ZiriTadi patroni-cxovelidan ara ZiriTad

patronze_praqtikulad ZuZumwovarTa TiTqmis yvela saxeobis

dainficirebiT: garemo areSi daavadebis gamomwvevi mikrobis

sakmao gamZleobiT da maTi TviTgadarCenis didi unariT; agreTve

maRali virulentobiT, rac uzrunvelyofs organizmis ueWvel

dainficirebas da infeqciuri procesis ganviTarebas.

T. SamaTavas monacemebiT 1962-1980 wlebSi saqarTveloSi

arakeTilsaimedo raionebis ricxvi 38,4-73,9%-is farglebSia.

epizootiurobis indeqsi (arakeTilsaimedo wlebis Sefardeba

8

dakvirvebis periodTan) yvelaze maRalia samaCablos, duSeTis,

goris, mcxeTis, gardabnis, adigenis, bogdanovkis, bolnisis,

TeTri wyaros, walkisa da siRnaRis raionebSi. es maCvenebeli

yvelaze dabalia onis, tyibulis, WiaTuris da Terjolis

raionebSi.

amJamindeli monacemebiT saqarTveloSi brucelozi yvela

regionSia fiqsirebuli, gansakuTrebiT aRmosavleT

saqarTveloSi, rac upirveles yovlisa, dakavSirebulia

pirutyvis Senaxvis sistemasTan. avadmyofoba yvelaze metad

gvxvdeba iq sadac pirutyvis momTabare Senaxvaa. maT

teritoriaze gadis pirutyvis gadasareki trasebi an

teritoriebi, romlebic momTabare mecxoveleobis raionebis mier

sazafxulo saZovrebadaa gamoyenebuli, sazafxulo saZovrebze

pirutyvis gadarekvisas yovelTvis ar icaven veterinarul-

sanitariul wesebs, amitom brucelozze arakeTilsaimedo

meurneobis pirutyvi, (sadac imyofeba avadmyofi cxovelebi),

ainficirebs gadasarek trasas, wyals, sazafxulo saZovars. misi

meSveobiT avaddeba keTilsaimedo meurneobis pirutyvic. (j.

babakiSvili Tanaavt. 2005w.) avadobis xvedriTi wili maRalia

gardabnis, dedofliswyaros, siRnaRis, dmanisisa da kaspis

raionebSi. aRniSnuli maCvenebeli yvelaze mcirea WiaTuris,

tyibulis, onis, da saCxeris raionebSi.

saqarTveloSi dRes-dReobiT ar tardeba brucelozis

winaaRmdeg gegmiuri, mizanmimarTuli daQqmediTi RonisZiebebi,

droulad ar xdeba daavadebuli cxovelis gamovlineba da misi

izolireba, rac warmoSobs brucelozis farTed gavrcelebis

saSiSroebas cxovelebSi da adamianebSi.

bolo periodSi bruceloziT daavadebis SemTxvevebi

TbilisSi mniSvnelovnad gaizarda. amis mizezia vet. sanitaruli

9

kontrolis dabali done, ris gamoc savaWro qselSi xvdeba

brucelozis aRmZvreliT dainficirebuli produqtebi, maT

Soris rZe da rZis produqtebi, romlebic aris daavadebis

gavrcelebis ZiriTadi wyaro. aRsaniSnavia is faqti, rom furis

jiqanSi brucelebi SeiZleba 7-9 wlamde arsebobdnen.

infeqciis droulad diagnostireba brucelozis enzootiis

prevenciisa da epizootiuri situaciis aRmofxvris yvelaze

saimedo da efeqturi meTodia.

kvlevis mizani. brucelozis problemidan gamomdinare

sadisertacio naSromis mizania:

1. saqarTveloSi msxvilfexa pirutyvis brucelozze

epizootiuri situaciis Seswavla.

2. Tanamedrove serologiuri reaqciebiT cxovelTa sisxlis

Sratebis gamokvleva.

3. serologiuri reaqciebis roz-bengal sinjis da konkurentuli

imunofermentuli analizis sadiagnostiko mniSvnelobis

SedarebiTi Seswavla.

4. msxvilfexa pirutyvidan gamoyofili Br. abortus bovis genetikuri

analizi da sxva saxeobis sasoflo-sameurneo cxovelebSi

migraciis dadgena.

kvlevis ZiriTadi amocanebi:

1. saqarTveloSi msxvilfexa pirutyvis brucelozze

epizootologiuri situaciis Seswavla.

2. msxvilfexa pirutyvis sisxlis Sratebis gamokvleva roz-

bengal sinjiT da imunofermentuli analiziT.

10

3. furebis rZis brucelozze rgoluri reaqciiT gamokvleva.

4. msxvilfexa pirutyvis paT. masalidan (sisxli, rZe, mogdebuli

nayofi) gamoyofili kulturebis myisieri polimerazuli

jaWvuri reaqciiT gamokvleva (RT-PCR).

5. AMOS-PCR-is gamoyeneba Brucella-s saxeobaTa

idetificirebisaTvis.

kvlevis siaxle: _ pirvelad saqarTveloSi msxvilfexa

pirutyvSi brucelozis diagnostirebisaTvis dRemde arsebuli

aglutinaciisa da komplementis SeboWvis reaqciis nacvlad

rekomendebulia maRalmgrZnibiare, swrafi da maRalefeqturi

kvlevis _ meTodebis konkurentuli imunofermentuli analizi

da polimerazuli jaWvuri reaqciis gamoyeneba, AMOS-PCR-iT

brucelas saxeobaTa diferencireba.

disertaciis praqtikuli mniSvneloba:

_ daixveweba brucelozze msxvilfexa pirutyvis serologiuri

meTodiT gamokvleva.

_ Seiqmneba sruli informacia genetikuri analiziT brucelebis

biotipebze, maT bunebaSi migraciaze.

_ miRebuli Sedegebis analizi safuZvlad daedeba brucelozTan

brZolis RonisZiebebis swor dagegmvasa da gatarebas.

kvlevis Sedegebis aprobacia:

naSromis ZiriTadi debulebebi moxseniebulia: a.i.p. saqarTvelos

agraruli universitetis:

a) doqtorantTa samecniero konferenciaze (2010 weli, ianvari)

11

b) amave universitetis doqtorantTa samecniero konferenciaze

(2010 weli, aprili)

v) infeqciur da invaziur sneulebaTa departamentis

saprezentacio komisiis sxdomaze (2010 weli, 10 maisi).

d) saerTaSoriso konferenciaze `agrobiomravalferovnebis

dacva soflis meurneobis mdgradi ganviTareba”, a.i.p. agraruli

universiteti (2010 weli, 24-25 noemberi).

e) doqtorantTa samecniero konferenciaze, a.i.p. agraruli

universiteti (2011weli. 15-19 marti).

kvlevi Sededgebis publikacia: Temis garSemo dargobriv

referirebul periodul gamocemebSi gamoqveynebulia 5

publikacia, maT Soris 3 individualuri avtorobiT, xolo ori

TanaavtorebTan erTad.

naSromis struqtura da moculoba: disertaciis teqsti moicavs

kompiuterze nabeWd 128 gverds da Sedgeba: Sesavlis,

literaturis mimoxilvis, sakuTari gamokvlevebis, miRebuli

Sedegebis ganxilvis, daskvnisa da praqtikuli winadadebebisagan.

sakuTari gamokvlevebi Sedgeba kvlevis obieqtebis, meTodebisa

da kvlevis Sedegebisagan. naSromi ilustrirebulia 11 cxriliT,

13 fotosuraTiT, 4 diagramiT, 1 ruqiT da 1 algoriTmiT.

sadisertacio naSroms Tan erTvis gamoyenebuli literaturis

CamonaTvali 213 publikacia.

12

2. literaturis mimoxilva

2.1. Bbrucelozi. istoria, gavrceleba sasoflo-sameurneo

cxovelebSi

Bbrucelozi miekuTvneba uZveles daavadebaTa ricxvs. adamianis

bruceloziT daavadebis pirveli Setyobineba gvxvdeba berZeni

eqimisa da filosofosis hipokratesa da aristoteles

naSromebSi. brucelozis Seswavlis istoriul etapad iTvleba

1886 weli, roca ingliselma samxedro eqimma devid brusma

kunZul maltaze mkvdari jarskacis elenTis paTologiuri

masalidan damzadebuli nacxebis mikroskopirebisas aRmoaCina

misi gamomwvevi mikrobi. Mmanve 1887 wels gamoyo aRmZvrelis

kultura romelsac uwoda maltis mikrokoki, xolo daavadebas

Sesabamisad maltis cxeleba. (CBRNE. 2001).

d. brusis mier istoriuli mniSvnelobis aRmoCenidan aTi

wlis Semdeg, ingliselma a. raitma da d. semplem (A.Wright,

D.Semple 1897w.) daadgines, rom bruceloziT daavadebuli

adamianis sisxlis Srats unari aqvs moaxdinos maltis

mikrokokis kulturis aglutinacia, aRniSnuli daedo safuZvlad

brucelozis serodiagnostikas. romelsac ewoda raitis reaqcia. veterinariisa da soflis meurneobis praqtikisaTvis didi xnis

winaT iyo cnobili Zroxebis Taviseburi daavadeba, romelic

make cxovelebSi iwvevda ganuviTarebeli nayofis mogdebas, anu

e.w. msxvilfexa pirutyvis infeqciuri-epizootiuri aborti. 1897

wels danielma eqimma da veterinarma bernard bangma da v.

striboltma (B. Bangand, V. Stribolt 1897) Seiswavles da daadgines,

rom Zroxebis infeqciuri abortis aRmZvreli aris

13

gansakuTrebuli `bacila~ (ase uwodes). mikroorganizmi

mkvlevarebma pirvelebma aRmoaCines abortirebuli Zroxis

saSvilosnosa da sanayofe garsebs Soris eqsudatSi, aRmZvreli

gamoyves sufTa mikrobuli kulturis saxiT da uwodes Bacillus

abortus bovis (Bang).

1916-1918 wlebSi, amerikelma mkvlevarma qalma elis ivensma (A.

Evans) moaxdina Micrococcus melitensis-is da Bacterium abortus-is

mikrobuli kulturebis SedarebiTi Seswavla da daamtkica maT

Soris mniSvnelovani msgavseba. e. ivensis gamokvlevebiT,

mikrobTa orive saxeoba morfologiuri da kulturaluri

TvisebebiT ar gansxvavdebodnen erTmaneTisagan. maTi gansxvaveba

SesaZlebeli aRmoCnda aglutinaciis reaqciiT.

1914 wels j. traumma abortirebuli Roridan gamoyo

brucelas mesame saxeoba da uwoda _ B. abortus suis.

1920 wels, k. meierma da feziem (K.Meier and Feusier) samive

mikrobi, kerZod: Micrococcus Melitensis, Bacterium Abortus da B. abortus

suis. gaaerTianes erT jgufSi, saerTo saxelwodebiT brucela

(Brucella), -brusis sapativsacemod. (Н. М. Колычев, Р. Г. Росманов.,

2003г.)

1953 wels badli, swavlobda ra cxvris infeqciur

epididimits, gamoyo B. ovis. 1957 wels stoenergma da lekmanma,

buCqnaris virTxis leSidan gamoyves mikroorganizmi, romelic

brucelas identuri aRmoCnda da uwodes B. neotome. 1966 wels ki

k. maiklma gamoyo ZaRlebSi brucelas aRmZvreli da uwoda

B. canis.

brucelozi cxovelTa qronikulad mimdinare infeqciuri

14

daavadebaa.Gavaddeba adamianic. igi miekuTvneba

zooanTroponozebs.

bruceloziT avaddeba yvela saxeobis Sinauri cxoveli, magram

ufro metad msxvilfexa pirutyvi, cxvari, Txa, Rori. brucelozi

dadgenilia mraval gareul, gansakuTrebiT balaxismWamel

cxovelSi, inficirdebian frinvelebic. aRwerilia brucelebis

civsisxlianTa organizmidan gamoyofis SemTxvevebi.Bbrucelozis

aRmZvreli gamoyofilia mRrRnelebidan, sisxlmwovi

fexsaxsrianebidan, eqtoparazitebidan. laboratoriuli

cxovelebidan yvelaze metad amTvisebelia zRvis goWi da TeTri

Tagvi. (j. babakiSvili, Tanaavt. 2005w.). brucelozis

mimarTAaxalgazrda cxoveli ufro gamZlea sqesobrivad

momwifebulTan SedarebiT.

brucelozze amTvisebelia xerxemlian cxovelTa 60 saxeobaze

meti, brucelebi gamoyofilia sisxlmwovi tkipebis 30 saxeobidan,

rwyilis ori saxeobidan, agreTve koRos ori saxeobidan da

saxlis buzebidan. brucelebi aRmoCenilia CrdiloeTis irmis

kanqveSa tkipas larvebSi.

aRsaniSnavia rom, infeqciis pirvelwyaros warmoadgenen ara

gareuli, aramed sasoflo sameurneo cxovelebi. daavadebis

erTeuli SemTxvevebi SeiZleba dafiqsirdes brucelozis

gadatanisas gareuli cxovelidan Sinaurze, Tumca daavadebis

saerTo epizootiur jaWvSi mniSvnelovan rols ar TamaSobs. (Т.

С. Костенко, Е. И. Скаршевская; 1989г.).

infeqciis aRmZvrelis wyaroa avadmyofi cxoveli. igi mik-

robs gamoyofs mogdebul nayofTan, sanayofe garsebTan da sana-

yofe siTxesTan, SardaTan, rZesTan da sxva gamonayofebTan er-

15

Tad. maTi meSveobiT inficirdeba sakvebi, wyali, sadgomi, movlis

sagnebi da sxva. (И. Ф. Таран, Г. И. Лямкин., 1996.)

brucelozi make cxovelebSi umTavresad mimdinareobs

abortebiT, mastitiT, saxsrebis anTebiT. mwarmoebelSi orqitiT

da epididimitiT. brucelozis gamavrcelebelia mdedr

pirutyvTan erTad mwarmoebelic (kerati, kuro, erkemali). Ffuri

rZesTan erTad brucelas gamoyofs xuT wlamde.

Bbrucelozis aRmZvrelis gadacema xdeba pirdapiri kontaqtiT

da dainficirebli garemo aris obieqtebis meSveobiT, pirveli

xorcildeba dagrilebis, xelovnuri daTesvlis (Tu mwarmoebeli

avadmyofia) dros.Bbrucelebi cxovelTa organizmSi SeiWreba

saWmlis momnelebeli sistemis, cxviris, Tvalis, sasqeso

organoebis lorwovani garsidan, aseve kanidan.

Aavadmyofobis gavrcelebaSi didi mniSvneloba aqvs

dainficirebuli rZis, moumwifebeli (axali, umarilo) yvelis,

brinzas, araJanis, karaqis, (A. Alim, Z. D. Tomul; 2005.), agreTve

sakvebad Termulad daumuSavebeli sakvebis miRebas (sisxliani

xorci, Tavis tvini da a.S.). (R. Hurtado. 2001). adamianidan adamianze

gadacema Zalian iSviaTia, arsebobs publikaciebi dedidan

nayofze daavadebis vertikaluri gadacemisa, amasTan ar aris

gamoricxuli sqesobrivi gziT aRmZvrelis gadacema. (CBRNE.

2001).

Aavadmyofobis gavrcelebaSi monawileobs adamiani, mecxove

leobis dargis muSaki, sasaklaoebisa da xorcis gadamamuSave-

beli punqtebis TanamSromlebi, romlebic ar icaven

veterinarul-sanitarul wesebs. (Z. Landau; L. Green., et. al. 1999;)

janmrTelobis dacvis saerTaSoriso organizaciisa da saerTa-

16

Soriso epizootiuri biuros monacemebiT, cxovelTa brucelozi

gvxvdeba mTels msoflioSi. SedarebiT farTodaa gavrcelebuli

msxvilfexa pirutyvis brucelozi.

17

2.2. brucelebis nairsaxeobebi, migracia cxovelebSi,

paTogenezi

brucelebi miekuTvnebian Schizomycete–s klass, rigi - Eubacteriale,

ojaxi – Brucelaceae. brucelebi aerobuli, katalaza da oqsidaza

dadebiTi, gramuaryofiTi, ujredSida fakultaturi

kokobacilebia. (R. Hurtado. 2001;) kafsulas ar warmoqmnian,

gamoimuSaveben ureazas. brucelebi warmoadgenen nitritebis

nitratebad gardaqmnis katalizators. (CBRNE.2001.). brucelas

kokovani formis zomebia 0,3-0,6 mkm-ia, xolo Cxirisebri formebis

0,6-2,5 mkm. genomis saSualo molekuluri masa 2,37X109 daltonia

(M. J. Korbet 1997.) brucelebi gamonaklisad warmoqmnian L-formebs.

brucelebs aqvT lipopolisaqariduli garsi, romelic sxva

gramuaryofiTi mikroorganizmebisagan gansxvavebiT naklebad

pirogenulia, rac ganapirobebs daavadebis dros maRali

cxelebis gamonaklisad gamovlinebas. (CBRNE.2001.).

lipopolisaqaridi (LPS) antigenia, romelic inducirebs

antisxeulebis gamomuSavebas. S-LPS (S-smooth –gluvi), gluvi

lipopolisaqariduli membrana savaraudod TamaSobs garkveul

rols mikrobis ujredSida gadarCenaSi. amasTan gluvi formis

Stamebi ufro gadarCebian vidre xorkliani. sarwmuno monacemebiT

S-LPS warmoadgens virulentobis mTavar faqtors, Tumca

paTogenezSi misi roli ar aris dazustebuli, rac miuTiTebs

ara ujredSida, aramed ujredgare gadarCenis rolze. (J. E. Ugalde,

C. Czibener, et. al. 2000).

brucelas ujredis kedelis struqturaSi Sedis Rrma O-antigeni

18

_ cilis kompleqsi, da zedapirul – S antigeni- lipiduri

kolmpleqsi. am ukanasknelis ori varianti arsebobs _ A da M

anu somaturi antigenebi A ( abortus) da M. (melitensis).

B. abortus – Stamebi Seicavs didi raodenobiT A antigens. kerZod A

da M antigenis Tanafardoba B. abortus-Si Seadgens 20:1. fiqroben,

rom es antigenebi warmodgenilia rTuli glucido-

lipidopolipeptiduri kompleqsiT. isini aqtiurad axdenen

antigenis sinTezis inducirebas, precipitaciis, aglutinaciis

k.f.r, da sxva reaqciis gamovlinebisas.

amJamad brucelas 7 saxeobaa cnobili: (cxrili 1). antigenuri

da bioqimiuri Tvisebebis mixedviT zogierT saxeobas aqvs

biovarebi.

1) B. abortus- msxvilfexa pirutyvis (aqlemis, iakis da kameCis

CaTvliT) brucelozis aRmZvreli, igi adamianis paTogenicaa. is

aerTianebs 9 biovars (1; 2; 3; 4; 5; 6; 7; 8 da 9), B. abortus zogierTi

biotipi (mag. 3, 6, 7 da 9) SedarebiT maRali virulentobis gamo,

romelic TiTqmis B. melitensis-is virulentobis msgavsia, iwveven

brucelozis epidemias mZime klinikuri formiT.

2) B. melitensis- cxvris da Txis brucelozis aRmZvreli; aqvs 3

biovari (1; 2; 3;). Aavaddeba adamianic.

3) B. suis- Roris brucelozis aRmZvreli; aqvs 4 biovari (1; 2; 3; 4;)

avaddeba adamianic. maT Soris biovari 1 da 3 iwvevs brucelozs

RorebSi; biovari 2 -kurdRlebSi da adamianSi, biovari 4

asnebovnebs CrdiloeTisa da kanadur irmebs.

4) B. canis – biovari ar gaaCnia. avaddeba ZaRlebi da misi msgavsi

19

cxovelebi, romlebmac SeiZleba daavadon adamianic.

5) B. ovis- erkemlis epididimitis aRmZvreli, avaddeba cxvari, igi

ar iwvevs adamianis daavadebas.

6) B. neotomae- buCqnaris virTxis brucelozis aRmZvreli, adamiani

ar avaddeba.

7) B. maris zRvis ZuZumwovrebis brucelozis aRmZvrelis Stamebis

formaluri saxelia, romelic Tavis mxriv kidev iyofa or

qvesaxeobad: 1) B. pinnipedialis-avaddeba fexfarflianebi: selapi,

zRvis lomi, da lomveSapi. 2) B. ceti, romelic gamoyofilia

veSapisebrTa ojaxis warmomadgenlebidan; (veSapi, delfini da

zRvis Rorebi). yvela es Stami gamoyofil iqna a.S.S-Si da

gaerTianebul samefoSi zRvis ZuZumwovrebidan. (G. Foster et. al. 1996.,

L.B. Forbes, et.al. 2000., A.R. Spickler, J.A. Roth, et.al. 2010, A.Whatmore et .al.,

2011.)

aRsaniSnavia rom, brucelas es saxeoba aavadebs ara marto

xmeleTis ZuZumwovrebs, aramed adamiansac. (riskis qveS arian

delfinebisa da selapebis momvlelebi). (S.D. Brew, L. L. Perrett; et. al.

1999., A. Whatmore, C. Dawson, 2011.)

brucelas saxeobisaTvis damaxasiaTebelia SerCeviTi

paTogenoba Sesabamisi saxeobis cxovelebis mimarT, magram

aRiniSneba maTi migraciis farTo speqtric. Bbrucelebs Tavisi

ZiriTadi Sualeduri patronis organizmis garda SeuZliaT zrda-

ganviTareba da paTologiuri procesis gamowveva sxva saxeobis

cxovelebSi; ase magaliTad, Txisa da cxvris brucelozi aRmZvr-

eli aavadebs Zroxasac, Rorsac da piriqiT. adamianis daavadeba

20

cxrili 1

brucelozi sasoflo-sameurneo da gareul cxovelebSi

#

organizmi

rezervuari cxovelebi

Ggeografiuli gavrceleba

1. B. abortus Zroxa, bufalo, iaki, aqlemi mTels msoflioSi

2.

B. melitensis

Txa, cxvari, aqlemi

Sua azia, laTinuri amerika, afrikis

nawili, zogierTi samxreT evropis

qveynebi

3.

B. suis

Rorebi (biotipi 1-3)

samxreT amerika, samxreT azia, a.S.S.

4.

B. canis

ZaRlisebrni

_

5.

B.ovis

cxvari

Aadamianis daavadebis SemTxveva ar aris

cnobili

6. B. neotomae mRrRnelebi Aadamianis daavadebis SemTxveva ar aris

cnobili

7.

B. pinnipediae and

B. catacaeae

zRvis ZuZumwovrebi,

veSapi, delfini, selapi

Uuaxlesi monacemebi, sadac aRwerilia

adamianTa daavadebis SemTxvevebi

21

samive ZiriTadi saxeobis sasoflo-sameurneo cxovelis

brucelozis aRmZvrelebs (B. abortus, B. melitensis, B. suis) SeuZlia met-

naklebi siZlieriT.Aamrigad, brucelebi `xatovnad ar

daikargebian.~ aRmZvrelis es Tviseba kidev ufro amZimebs am

infeqciis saSiSroebas, adamianebsa da cxovelebSi. (m.

lomineiSvili, T. gavaSeli. 1995)

B brucelebi miekuTvnebian paTogenur mikrobTa jgufs.

paTogenobiT da virulentobiT brucelebis sxvadasxva saxeobebi

gansxvavebulia. aRniSnuli exeba ara marto sxvadasxva saxeobebs,

aramed erTi da igive saxeobebSi Semaval sxvadasxva

serovariantebs. (Н. Е. Ненатов 1990., В. М. Авилов 1997.)

Tanamedrove SexedulebiT, brucelozis paTologiaSi, arCeven,

cxovelTa dainficirebis, erTmaneTisagan gansxvavebul or

moments: pirveli cxovelis organizmSi infeqciis gamomwvevi

mikrobebis mcire dozebiT TandaTnobiTi dagroveba, infeqciuri

gamRizianeblis sumacia, romelic infeqciuri procesis sawyis

etapze mimdinareobs imunuri sistemis gaRizianebis _ aRgznebis

gareSe. meore - cxovelis organizmSi aRmZvrelis didi

raodenobiT erTdrouli moxvedra, rasac mosdevs imunuri

gardaqmnis (Zvrebis) gamovlineba dainficirebidan pirvel

kviraSi.

veterinarul praqtikaSi gansakuTrebuli mniSvnloba aqvs

pirvel moments, vinaidan am dros ar mJRavndeba imunuri

gardaqmnebi dainficirebul organizmSi. sasoflo-sameurneo cxo-

velebi, rogorc wesi, imyofebian umTavresad jgufuri Senaxvis

22

pirobebSi, sadac fiziologiuri TvalsazrisiT mkveTrad

gansxvavebuli individebia Tavmoyrili, romlebic brucelozis

amTviseblobasTan damokidebulebaSi amJRavneben metnakleb

mgrZnobelobas. cxovelebs Sors xdeba mudmivi kontaqti,

rogorc pirdapiri, aseve arapirdapiri gziT. amdenad infeqciis

keris gaCenisas cxovelebs Soris warmoiSveba dainficirebisa da

daavadebis gamovlenis sxvadasxva pirobebi. swored aRniSnuli

gansxvavdeba cxovelTa dainficirebis savele pirobebi

eqsperimentulisagan. eqsperimentSi dadgenilia brucelebis

miaxloebiTi dozebi, romlebic iwveven mxolod regionaluri

limfuri kvanZebis inficirebas e.w.- regionaluri infeqcia da

septikur process-generalizebuli infeqcia. msxvilfexa

pirutyvisaTvis saSualo damainficirebeli doza meryeobs B.

abortus-is 10-dan 100 aTas cocxali mikrobuli ujredis

farglebSi. cxovelis mier ufro maRali dozebis miRebis

SemTxvevaSi viTardeba Zlieri da intensiurad gamoxatuli

infeqcia, imunologiuri reaqtivobis gamomJRavnebiT,

dainficirebidan SedarebiT mokle periodSi. dadgenilia, rom

zogierTi cxovelis organizmSi ganmeorebiT brucelebis

subminimaluri raodenobis moxvedrisas mikrobebis sumaciis

Sedegad aRmocendeba tipiuri daavadeba.

bruceloziT dainficirebis pirveli gza meorisagan principSi

imiT gansxvavdeba, rom aRmZvrelis dabali dozebi, saSiSi ar

aris, magram erTi da igive cxovelis organizmSi misi mravaljer

moxvedrisas brucelebi mravldebian, vrceldebian da iwveven

baqteriemiasa da klinikurad srulyofilad Camoyalibebul

23

brucelozs. am SemxvevaSi daavadebis gamowveva SeuZlia

erTjeradad damainficirebel dozaze naklebi raodenobis

mikrobebsac, e.i. ar aris aucilebeli sumaciis Sedegad

maqsimaluri damainficirebeli dozis dagroveba. amdenad

araimunuri cxovelis organizmSi, iqmneba faruli

brucelamtareblobis pirobebi.

Bbrucelozis aRmZvrelebi sakmaod `agresiulebi~ arian da

xasiaTdebian maRali invaziurobiT. maT SeuZliaT organizmSi

SeRweva dauzianebeli lorwovani garsebidanac. amasTan isini

amJRavneben hemotoqsiurobas. brucelebi miekuTvneba ujredSida

parazitebs. isini cocxloben da mravldebian retikulo-

endoTeliaruli sistemis ujredebSi, amasTan maT SeuZliaT

agreTve ujredgare parazitobac. (М. М. Ременцова, 1988., И. Ф. Таран,

Г. И. Лямкин 1996.)

organizmSi SeWrili brucelebi garkveuli drois ganmavlobaSi

axlomdebare limfur kvanZebSi ibudeben da mravldebian.

organizmis winaaRdegobis daZlevis SemTxvevaSi, isini gadadian

Sinagan organoebSi, infeqcia Rebulobs generalizebul formas.

3-4 da meti kviris Semdeg avadmyofoba gadadis latentur

formaSi, brucelebis curSi an calkeul limfur kvanZSi

lokalizaciiT. Bbrucelebi gamravlebisaTvis yvelaze

xelsayrel garemos make cxovelis saSvilosnoSi pouloben,

sadac isini didi raodenobiT mravldebian, iwveven saSvilosnosa

da sanayofe garsebis anTebiT movlenebs, ris Sedegadac irRveva

kavSiri dedis organizmsa da nayofs Soris, es ukanaskneli

kvdeba da rogorc organizmisaTvis ucxo sxeuli gareT

24

gamoidevneba an cocxali, dedis organizms scildeba

saSvilosnos nervuli SekumSvis Sedegad, nayofis mogdebis

Semdeg brucelebi umTavresad cursa da limfur kvanZebSi

Caibudeben. organizmSi irRveva nivTierebaTa cvla, rac

gamovlindeba sisxlis maragis tutianobis, kalciumis

SemcirebiT da araorganuli fosforis momatebiT. make cxovelis

saSvilosnoSi gamoiyofa eriTritoli, romelic axdens

brucelebis gamravlebis stimulirebas. amrigad, brucelozis

CamoyalibebaSi sami faza aRiniSneba _ pirveladi latencia

(regionaluri infeqcia), generalizacia da ganmeorebiTi

latencia. (j. babakiSvili Tanaavt. 2005w).

25

2.3.B veterinariasa da medicinaSi brucelozis serologiuri

da genetikuri diagnostika

infeqciur daavadebaTa sadiagnostikod veterinarul

praqtikaSi gamoiyeneba mikroskopuli, baqteriologiuri,

bioqimiuri, serologiuri meTodebi. (Б. И. Антонов 1986., Н. Р.

Ассонов 2002., Н. М. Колычев с соавт. 2003., J. B. Muma, et. al. 2007.) es

ukanaskneli damyarebulia imunologiur reaqciebze. am movlenas

safuZvlad udevs antigenisa da specifikuri antisxeulis

Semcveli Sratis urTierTqmedeba.

imunologiuri reaqciebi maRalmgrZnobiare da specifikuria.

(Э. М. Плотникова 2001., Э.М. Плотникова с соавт. 2010.) sadiagnostikod

gamoyenebul serologiur reaqciaSi erT-erTi komponenti unda

iyos cnobili. misi specifiuroba saSualebas gvaZlevs

davadginoT meore – ucnobi komponentis arseboba.

imunologiuri reaqciebis msvlelobaSi ganarCeven or

urTierTdakavSirebul da Tanamimdevrul fazebs: pirvel fazaSi

antisxeulebi ganicdian adsorbcias antigenze. es faza TvaliT

uxilavia, vinaidan mimdinareobs garegnulad SesamCnevi efeqtis

gareSe. meore faza, romelic koloiduri procesis analogia,

xasiaTdeba antigenis lizisiT, SewebebiT, daleqviT da sxva, rac

Sedegs xiluls xdis.

serologiuri reaqciebiT specifiuri antisxeulebis aRmoCena

gacilebiT xelmisawvdomia vidre aRmZvrlis gamoyofa. mTel rig

SemTxvevaSi, dadebiTi serologiuri reaqciebi cxovelTa

organizmsa da Sesabamisi infeqciis aRmZvrelis urTierTqmedebis

dadgenis utyuari saSualebaa. serologiuri reaqciiT

26

SesaZlebelia klinikurad msgavsi niSnebiT mimdinare infeqciur

daavadebaTa diferenciacia. (Х. Х. Абдулин 1980.,)

praqtikaSi, serologiuri reaqciebi warmatebiT gamoiyeneba

infeqciis afeTqebis da infeqciuri daavadebis wyaros

dasadgenad, infeqciuri daavadebis sadiagnostikod mowodebulia

aglutinaciis (modifikaciebiT), precipitaciis, komplementis

fiqsaciis, neitralizaciis, da sxva. (m. naTiZe Tanaavt. 2012w.,

Р.А. Нуратинов, 1993.) reaqciebi.

aglutinaciis (raitis) reaqci (ar): imunuri SratiT (marilTa

xsnarebis TandaswrebiT) mikrobuli an sxva warmoSobis ujredis

(eriTrocitebi da sxva) Sewebebas aglutinacia ewodeba.

es testi pirvelad iqna SemoTavazebuli 1897 wels raitisa da

semples mier adamianebSi maltis cxelebis sadiadnostikod.

1909 wels qristensenma da 1910 wels holtma es reaqcia dadges

msxvlfexa pirutyvSi infeqciuri abortis sadiagnostikod.

aglutinaciis reaqcia gamoiyeneba brucelozis, botulizmis,

cxenis infeqciuri abortis, salmonelozis da sxva infeqciuri

daavadebis sadagnostikod.

aglutinaciis reaqciis dadebiTi mxarea is rom, is iZleva

dadebiT Sedegebs daavadebis adreul stadiaSi, magram aris

iseTi SemTxvevebic rodesac aglutinaciis reaqciis uaryofiTi

Sedegi ar gamoricxavs infeqciis arsebobas. meore mxriv man

SeiZleba mogvces dadebiTi Sedegi iseT naxirSi romlebic

iTvleba brucelozze keTilsaimedod. aglutinaciis reaqcias

dgamen sxvadsaxva meTodiT:

27

wveTovani anu firfitovani aglutinacia, gamoiyeneba cxovelis

swrafi gamokvlevisaTvis.

mikroskopuli aglutinaciis modificirebuli meTodi farTod

gamoiyeneba leptospirozis sadiagnostikod.

sisxlwveTovani aglutinacia gamoiyeneba pulorozze frinvelis

masobrivi gamokvlevisaTvis.

makroskopuli aglutinacia anu moculobiTi aglutinacia:

aglutinaciis reaqciis mgrZnobelobis da specifikurobis

gazrdisaTvis ixmareba erTiani mSrali brucelozuri antigeni.

(gamoiyeneba aglutinaciis reaqciasa da komplementis fiqsaciis

reaqciaSi). am meTodiT xdeba brucelozis gamokvleva. (m.

TavamaiSvili 1988w.)

xedlsonis aglutinaciis reaqcia: warmatebiT gamoiyeneba

medicinaSi. xedlsonis reaqcia brucelozis diagnostikis

swrafi, specifikuri da maRalmgrZnobiare meTodia. (m. naTiZe

Tanaavt. 2005w., m. naTiZe, m. sarjvelaZe 2006w.) adamianebSi

brucelozis sadiagnostikod reaqciis ori varianti gamoiyeneba,

idgmeba moculobiTi da minaze _ swrafi meTodiT. am

ukanaskneli meTodiT reaqciis dadgma rekomendebulia mxolod

stacionaris pirobebSi donorebis masobrivi gamokvlevis dros.

aglutininebis titris da misi dinamikis gansazRvrisaTvis

aucilebelia moculobiTi meTodebis gamoyeneba.

arapirdapiri hemaglutinaciis reaqcia: maRalspecifikuri da

mgrZnobiarea. es reaqcia farTod gamoiyeneba mTeli rigi

infeqciuri daavadebebis, maT Soris jilexis aRmZvrelis sporo-

28

vani da vegetatiuri formebis, brucelebis aRmosaCenad da

identifikaciisaTvis, aRniSnuli reaqcia farTed gamoiyeneba

virusul daavadebaTa sadiagnostikod, maT Soris Turqulis

dros. reaqciaSi antisxeuluri eriTrocituli diagnostikumi

gamoiyeneba.

arapirdapiri hemaglutinaciis reaqcia mgrZnobiarea, misi

daxmarebiT SeiZleba sisxlis SratSi umciresi raodenobis

antisxeulebis aRmoCena. reaqcias safuZvlad udevs

eriTrocitebze antigenis adsorbcia da maTi Semdgomi

hemaglutinacia antigenis homologiuri antisxeulebiT.

komplementis fiqsaciis reaqcia (kfr). es aris rTuli,

maRalmgrZnobiare da specifikuri serologiuri reaqcia,

romelic didi xania gamoiyeneba cxovelTa brucelozis

sadiagnostikod. (J. Huber et. al. 1986., S. Sutherlandat. et. al. 1986.)

komplementis fiqsaciis reaqcia pirvelad gamoyenebul iyo 1909

wels msxvilfexa pirutyvis infeqciuri abortis

diagnostirebisas. is baqteriolizuri (antigeni, gamosakvlevi

Srati, komplementi) da hemolizuri (cxvris eriTrocitebi da

hemolizuri Srati) sistemebisagan Sedgeba. es reaqcia

saSualebas iZleva gamoavlindes brucelozis rogorc adreuli

aseve qronikuli formebi. igi ufro specifiuri da mgrZnobiarea

vidre aglutinaciis reaqcia, magram ufro rTulia da misi

gamoyeneba savele pirobebSi SeuZlebelia. komplementis

fiqsaciis reaqcia veterinarul praqtikaSi gamoiyeneba

brucelozis, msxvilfexa pirutyvis peripnevmoniis,

tuberkulozis, cxenis dagrilebis daavadebis, qoTaos,

29

Turqulis da sxva infeqciuri daavadebTa sadiagnostikod. (m.

naTiZe, t. gabisonia, s. rigvava. 1996w; m. naTiZe, t. gabisonia, T.

naTiZe, s. rigvava. 2005w.)

zogierTi mecnieri cdilobda gaezarda k.f.r-iis mgrZnobeloba

msxvilfexa pirutyvis brucelozis diagnostirebisas, rac xels

Seuwyobda ufro swrafad gamovlinebuliyo naxirSi daavadebuli

pirutyvi da Semcirebuliyo gamojanmrTelebis periodi

arakeTilsaimedo meurneobebSi. (Р.А. Нуратинов с соавт. 1993.)

komplementis fiqsaciis reaqciis paralelurad warmatebiT

gamoiyeneba komplementis fiqsaciis gaxangrZlivebuli reaqcia,

romelic gamoirCeva zedmiwevniT maRalmgrZnobelobiT da

specifiurobiT.

imuno-fluorescenciis reaqcia: gamoiyeneba infeqciuri

daavadebis eqspresdiagnostikisaTvis medicinasa da

veterinariaSi.

es meTodi damyarebulia zogierTi organuli da araorganuli

nivTierebis naTebis Tvisebaze, ultraiisferi sxivebiT

zemoqmedebis Sedegad. imunoflurescenciis meTodi swrafi da

xSir SemTxvevaSi specifikuria. misi daxmarebiT SesaZlebelia

erTeuli mikrobis da virusis aRmoCena. (Х. Х. Абдулин. 1980.)

imunoflurescencia warmatebiT gamoiyeneba cofis, jilexis,

salmonelozis, wiTeli qaris, brucelozis, yvavilis, Roris

Wiris da sxva infeqciur daavadebaTa sadiagnostikod. (m. naTiZe

1989w.)

30

brucelozi kacobriobisaTvis erT-erT ZiriTad problemad

rCeba. daavadebis faruli mimdinareoba arTulebs dadebiTad

moreagire cxovelebis gamovlinebas. avadmyofi-

baqteriamtarebeli cxoveli warmoadgens aRmZvrelis ZiriTad

rezervuars, rac qmnis janmrTel cxovelsa da adamianebSi infeq-

ciis gavrcelebis saSiSroebas.

cxovelTa brucelozis diagnostika, gansakuTrebiT

adreuli, infeqciis profilaqtikis safuZvelTa-safuZvelia.

bruceloziT daavadebuli cxovelebis drouli gamovlineba

rTulia, vinaidan organizmSi dainficirebis momentidan da

sisxlSi specifikuri antisxeulebis gamovlinebamde gadis didi

dro. aRsaniSnavia is garemoebac, rom antisxeulebis

gamomuSaveba da gadasvla sisxlis mimoqcevis sistemaSi ar

xdeba, konkretuli da zustad dadgenili periodis gavlis

Semdeg. Aam periodebs Soris intervali Zalian didia da

damokidebulia mraval faqtorze: mikrobis dozaze, misi

paTogenobis xarisxze, anu virulentobaze, cxovelis organizmis

individualur mrZnobelobaze, saerTo fiziologiur

mdgobareobaze, zogad rezistentobaze da sxva. zogierTi

cxoveli serologiuri testebiT gamokvlevisas, daificirebidan

ukve meSvide dReze iZleva dadebiT reaqcias. xSiria SemTxvevebi,

roca antisxeulebis diagnostireba, rigi cxovelebis organizmSi

aRmZvrelis moxvedridan, mxolod ramdenime Tvis Semdeg xdeba

SesaZlebeli.

brucelozis dros, sisxlis SratSi specifikuri antisxeulebis

31

gamoCena xdeba organizmSi aRmZvrelis SeWridan mxolod

garkveuli drois Semdeg da SenarCundebian xangrZlivi drois

ganmavlobaSi. msxvilfexa pirutyvSi aglutininebi gamovlindeba

cxovelis dainficirebidan 5-10 dRis Semdeg. aglutininebis

dagrovebis intensivoba, maTi titris done infeqciis

ganviTarebis procesSi meryeobs Zalian maRali ganzavebidan

srul gaqrobamde. rigi avtorebisa Tvlis, rom aglutininebis

dagroveba organizmSi dakavSirebulia da emTxveva infeqciis

aRZvris process da aqedan gamomdinare mikrobebis intensiurad

gamoyofas garemoSi. sayuradReboa Semdegi garemoebac:

aglutinaciis reaqciis ,,gamovardna” (gaqroba) SeiZleba moxdes

brucelozuri procesis gamwvavebis SemTxvevaSi. ase magaliTad

aRwerilia brucelozuri abortebis SemTxvevebi msxvilfexa

pirutyvSi uaryofiTi aglutinaciis reaqciis dros.

cxovelis sicocxleSi brucelozis diagnostika serologiuri

reaqciebiT xorcieldeba. (Р. А. Нуратинов, с соавт. 1993) maTi

saSualebiT SesaZlebelia dadgindes adamianisa da cxovelis

organizmebis paTogenTan kontaqti da daisvas diagnozi;

dReisaTvis brucelozis serologiuri diagnostikisaTvis

veterinarul praqtikaSi danergilia roz-bengal sinji, roz-

bengal testiT dadebiTi da saeWvo sisxlis Sratebi

dadasturebas saWiroebs serologiuri kvlevis Tanamedrove

meTodiT imunofermentuli analiziT. (D. Rylatt, et. al. 1985., О. Д.

Скляров., 2005., М. М. Желудков с соавт. 2003.).

roz-bengalis reaqcia (rbr): anu aglutinaciis reaqcia SratSi

32

bengalis vardisferi antigenis gamoyenebiT, es aris swrafi

reaqcia, romelic SemuSavebuli iqna amerikeli mecnierebis mier.

(W. Morganat. et. al. 1969., J. Huber et. al. 1986.) antigeni warmoadgens 8%-

ian B. abortus Stami 19-is mokluli ujredebis suspenzias,

buferul xsnarSi. xsnaris pH=3,65-ia (mJave are), romelic

SeRebilia bengalis vardisferiT. dReisaTvis es reaqcia

farTod gamoiyeneba msoflios bevr qveyanaSi (safrandeTi,

germania, aSS, inglisi da sxva). (M J. Corbel 1972). mas axasiaTebs

gansakuTrebulad maRali specifikuroba. (М. М. Иванов с соавт.1976.)

reaqciis dadgmis teqnika sakmaod martivia da SeiZleba Catardes

nebismier pirobebSi. (m. naTiZe Tanaavt. 2005w., W. Morgan, et. al. 1969.,

В. Б. Бельченко, С. Е. Сайдашева 1980.)

roz-bengal testiT xdeba B. abortus specifiuri antisxeulebis

aRmoCena adamianis, Sinaur da gareul cxovelebSi. cnobilia,

rom am skrining-tests aqvs 63 – 97% mgrZnobeloba da 70 – 99%

specifikuroba.

roz-bengal testi saSualebas iZleva dadgindesBbrucelebis

saxeobebis O-polisaqaridebis (OPS) mimarT aglutinirebadi

antisxeulebi. aRniSnuli mikroorganizmi zrdisaTvis saWiroebs

specialur garemos; aseTi garemo adamianebi da cxovelebia,

romlebic inficirebuli arian brucelebiT. (B. abortus, B. melitensis,

B. suis). dainficirebuli adamianisa da cxovelis organizmSi

brucelebi inducireben aglutininebis gamomuSavebas. sisxlis

OPS-specifiuri antisxeulebi jvaredinad emagrebian roz-benga-

lis antigens da iwveven mikroskopul aglutinacias.

33

Kkonkurentuli enzimuri imunofermentuli analizi: brucelozis

adreuli da zusti diagnostireba, warmatebuli mkunalobisa da

profilaqtikis sawindaria. erT-erTi aseTi serologiuri

meTodia imunofermentuli analizi, romelic maRalmgrZnobiare

da specifikuria. es meTodi warmatebiT gamoiyeneba medicinasa

veterinariaSi. (А. Я. Самуйленко, 2001.,). sxva msgavs meTodebTan

SedarebiT, rogoricaa imunofluorescentuli analizi, romlis

Sedegis Sefasebac xdeba subieqturad, xolo radiomunologiuri

meTodis gamoyenebisas ki iyeneben mokle periodis

naxevradgaxleCvad izotopebs, romlebic potenciurad saSiSia

adamianisaTvis, imunofermentul analizs es uaryofiTi mxareebi

ara aqvs, amitom misi gamoyeneba optimaluria. (Л. А. Тарасишин

1986., М. М. Желудков с соавт. 2003.)

imunofermentuli analizi farTod gamoiyeneba praqtikulad

yvela virusuli da baqteriuli daavadebebis sadiagostikod,

aseve gamoiyeneba parazituli, sokovani, onkologiuri

daavadebebis gamosakvlevad. (К. В. Шумилов с соавт. 2001., М. В.

Баландина с соавт. 2004., А. И. Федоров с соавт. 2007.)

jandacvis saerTaSoriso organizaciis mier,

imunofermentuli analizi CarTulia, rogorc aucilebeli

eqspres-diagnostikis testi, rogorc epizootologiuri aseve

laboratoriul kvlevaSi. (М. М. Желудков с соавт. 1989., Д. А.

Девришев с соавт. 2007.)

cnobilia imunofermentuli analizis ramdenime meTodi:

pirdapiri, arapirdapiri, `sendviCis~ tipis, mablokirebeli da

34

konkurentuli. yvelaze metad gamoiyeneba konkurentuli

imunofermentuli analizi.

Tanamedrove veterinarul praqtikaSi brucelozis

sadiagnostikod warmatebiT gamoiyeneba konkurentuli

imunofermenuli analizi, (C-ELISA) Brucella abortus da melitensis-is

sawinaaRmdego antisxeulebis aRmosaCenad Sinaur da gareul

cxovelebSi. (E. Engvall, P. Perlman 1997., J. W. Byrd, et. al. 1979., D. Gall, K.

Nielsen 1994., L. Samartinoet et. al. 1999., J. Paweskaet. et. al. 2002., K. Nielsenet

et. al. 2008.)Mmsxvilfexa pirutyvSi am analizis saSualebiT

SesaZlebelia, erTmaneTisagan gavarCioT Brucella-Ti

inficirebuli cxovelebi, Stami 19-iT vaqcinirebuli cxovelebi

da jvaredinad moreagire gramuaryofiTi baqteriebiT

inficirebuli cxovelebi. (C. Robleset. et. al. 2009.)

rZis rgoluri reaqcia: veterinariaSi brucelozis

sadiagnostikod aprobirebuli da farTed gamoyenebulia

furebis rZis gamokvleva rgoluri reaqciiT. aRniSnuli

reaqciaAASemoTavazebulia 1937 wels. rgoluri reaqciis

dasadgmelad SemuSavebulia feradi antigeni. (П. А. Триленко, 1954.,

Ф. С. Логинов, 1956., WJ. McCaughey 1972., J. Huberat. et. al. 1986., L. Sutraet.

et. al. 1986. )

Ereaqciis arsi imaSi mdgomareobs, rom rZeSi specifikuri aglu-

tininebis arsebobisas xdeba SeRebili antigenis Sejgufeba rac

warmoqmnis aglutinats, es ukanaskneli adsorbirdeba naRebTan

(rZis cximis burTulakebTan), maTTan erTad sinjaraSi

amotivtivdeba siTxis (rZisa da antigenis narevis) zedapirze

35

sadac warmoqmnis ferad rgols. rZis danarCeni masa

uferuldeba. uaryofiTi reaqciis SemTxvevaSi cximi (naRebi)

fers ar icvlis da rZe rCeba Tanabrad Seferadebul

mdgomareobaSi.

reaqciis dasadgmelad gamoiyeneba biofabrikaSi specialurad

damzadebuli antigeni, romelic brucelebis standartuli

Senawonia. antigeni hematoqsiliniT lurj an trifenol-

tetrazoliT an difenil-tetrazoliT muq-wiTel ferSi

SeRebili suspenziaa. Aantigeni mzaddeba B. abortus kulturidan,

romelic Camosxmulia flakonebSi. (О. И. Морякова, 1961).Aantigenis

Senaxvisas warmoiqmneba naleqi. xmarebis win SenjRrevisas is

advilad gadadis homogenur masaSi.

polimerazuli jaWvuri reaqcia: polimerazuli jaWvuri reaqcia

axali mZlavri meTodia, romelic saSualebas iZleva d.n.m-is

erTi aslisagan usazRvro raodenobis d.n.m-is aslebis

warmoqmnas. (M. Da Costa et. al. 1996.) idea ekuTvnis kari muliss 1980

wels da mas uwoda `polimerazuli jaWvuri reaqcia“ (PCR). anu

enzimuri amplifikaciis reaqcia in vitro. (K. B. Mullis, et. al. 1986.) am

meTodma revolucia moaxdina genetikur kvlevebSi, maT Soris

genetikuri daavadebebis diagnostikaSi, sasamarTlo medicinaSi

da molekuluri biologiis evoluciaSi. (M. Clementietal. 1993., И. Г.

Дубинина, 1998., С.В. Дентовская с соавт. 1999.)

ujredis daSlisas, dnm-is replikacia moicavs fermentebiT

ganpirobebul reaqciebs, romlis saboloo Sedegs warmoadgens

mTeli genomis zusti aslis Seqmna. fermentebi Tavdapirvelad

36

Slian dnm-is ormag jaWvs erTmag jaWvebad. Semdeg replikaciis

dasawyisSi, rnm polimeraza asinTezirebs mokle rnm. jaWvs,

romelic komplementuri iqneba dnm-is erT-erTi jaWvisa. es dnm-

rnm heterodupleqsi moqmedebs, rogorc dnm polimerazas

mimagrebis `praimingis~ adgili, romelic SemdegSi asinTezirebs

dnm komplementur jaWvebs.

1990 wels d. fekeTma (D. Fekete) moaxdina Brucella-as deteqcia

polimerazuli jaWvuri reaqciiT. (A. Fekete et. al. 1990.)

brucelebis indikacia da identifikacia sakmaod Sromatevadia,

iTxovs materialur danaxarjebs da dros.B

bolo aTwleulia, rac brucelozis sadiagnostikod

aqtiurad gamoiyeneba molekulur-biologiuri kvlevis

aRniSnuli meTodi. (С. В. Балахонов с соавт. 1996., О. Д. Скляров с соавт.

2005.)

polimerazuli jaWvuri reaqciiT SesaZlebelia Brucella-as

saxeobebis biotipebisa da Stamebis swrafi identifikacia da

diferencireba. (Ю. К. Кулаков с соавт. 2006., М. М. Желудков с соавт.

2007., B. J. Bricker 2002.,) polimerazuli jaWvuri reaqcia gamoyeneba

brcelozis sadiagnostikod da ufro mgrZnobiarea serologiur

testebTan SedarebiT. igi iZleva Brucella–is adreuli

diagnostikis saSualebas, kerZod, inficirebidan daaxloebiT,

aTi dRis Semdeg. Ppolimerazul jaWvur reaqciaze damyarebuli

meTodi asve saimedo aRmoCnda Brucella-is deteqciisaTvis

sxvadasxva nimuSSi, maT Soris Zroxis sisxlSi da rZeSi,

bunebrivad inficirebuli Zroxis organoebSi, kulturaSi da

37

sxva klinikur masalaSi, (C. Romero, et. al. 1995., А. Г. Яковлев с соавт.

2002.). igi saSualebas iZleva erTi samuSao dRis ganmavlobaSi

daadginos brucelas arseboba umniSvnelod kontaminirebul

nebismieri saxis nimuSSi. (К. В. Шумилов с соавт. 1996., И. Н.Шарова

2001., О. Д. Скляров с соавт. 2004.).

polimerazuli jaWvuri reaqciis meTodis meSveobiT

Teoriulad SesaZlebelia, sakvlev masalaSi aRmovaCinoT dnm-s

anu genetikuri masalis erTi asli anu meTodis mgrZnobelobas

zRvari ar aqvs. polimerazuli jaWvuri reaqciis erT-erTi

upiratesobaa absoluturi mgrZnobelobis paralelurad

specifikuroba. (D. S. Leal-Klevezas et. al. 1995., С. Б. Гаранина 1996., М. М.

Желудков с соавт. 2005.) gamokvlevis sworad Catarebis SemTxvevaSi

cru dadebiTi Sedegis miReba gamoricxulia. polimerazuli

jaWvuri reaqcia ar cvlis kvlevis tradiciul morfologiur,

bioqimiur an imunur meTodebs, is kvlevis aucilebel damatebiT

meTods warmoadgens.

AMOS PCR saSualebas iZleva movaxdinoT brucelas oTxi

saxeobis (A-abortus, M-melitensis, O-ovis, S-suis.) diferenciacia. (B. J.

Bricker, S. M. Halling 1994; B. J. Bricker. 2003.)

38

3. sakuTari gamokvlevebi

3.1. gamokvlevis masala da meTodebi

TemiT gaTvaliswinebuli samuSao Sesrulebulia 2008–2011w.w.Si.

kvleva mimdinareobda soflis meurneobis saministros

laboratoriaSi, S.p.s. ,,imunogenSi”.

cdebis msvlelobis procesSi gamokvlevas davuqvemdebareT:

2008-2011 w.w.-Si msxvilfexa pirutyvis 15175 sisxlis Srati, 97

rZis nimuSi. paralelurad 2010-2011 w.w. periodSi mimdinareobda

muSaoba msxvilfexa pirutyvis brucelozis aRmZvrelis

tipirebaze, am mizniT gamovikvlieT soflis meurneobis

saministros laboratoriaSi, saqarTvelos sami regionidan

(qvemo qarTli, kaxeTi da imereTi) Semosuli 1253 sisxlis 577

rZis nimuSi da 5 mogdebuli nayofi, agreTve 53 Sratis nimuSi.

sul gamovikvlieT 15228 sisxlis Srati, 1253 nativuri sisxli,

697 rZe da 5 mogdebuli nayofi. (cxrili 2); gamokvlevaTa

procesSi gamovyaviT da srul baqteriul analizs

davuqvemdebareT B. abortus 20 kultura.

gamokvleva mimdinareobda jandacvis saerTaSoriso

organizaciis (WHO) da cxovelTa janmrTelobis msoflio

organizaciis (OIE), mier dadgenili diagnostikuri testebiT,

romlebic gamoiyeneba rogorc epizootologiuri aseve

laboratoriuli kvlevisaTvis.

39

cxrili. 1

brucelozze gamokvleuli paT. masalis CamonaTvali da

raodenoba

#

weli

sisxlis

Srati

nativuri

sisxli

rZis

nimuSebi

Mmogdebu

li

nayofi

1.

2008

3082

280

106

_

2.

2009

6740

335

162

1

3.

2010

3654

450

237

2

4.

2011

1752

188

192

2

5.

sul

15228

1253

697

5

40

2008 wels Semovida da gamokvleul iqna 3082 msxvilfexa

pirutyvis sisxlis Srati, 280 nativuri sisxli da furebis 106

rZe.

2009 wels gamokvlevebs davuqvemdebareT msxvilfexa

pirutyvis 6740 sisxlis Srati, 335 nativuri sisxli, furebis 162

rZis nimuSi da 1 mogdebuli nayofi.

2010 wels gamokvlevebi Catarda msxvilfexa pirutyvis 3654

sisxlis Sratze, 455 nativur sisxlze, 237 rZesa da 2 mogdebul

nayofze.

2011 wels gamokvlevebi moicavda laboratoriaSi Semosul

1752 sisxlis Srats, 188 nativur sisxls, furebis 192 rZis nimuSs

da 2 mogdebul nayofs.

gamokvlevebisaTvis viyenebdiT baqteriologiuri da

imunologiuri kvlevis klasikur da Tanamedrove meTodebs.

mikrobTa kulturebi: kvlevisaTvis gamoyenebul iqna Cvens mier

cdebis procesSi gamoyofili da tipirebuli B. abortus 20

kultura. am mizniT SeviswavleT maTi morfologiur-

kulturaluri da zogierTi bioqimiuri Tvisebebi.

bioqimiuri aqtivoba: brucelebis bioqimiuri Tvisebebis dadgena

moicavda:

1) oqsidazas tests.

2) ureazas tests.

3) katalazas tests.

41

4) gogirdwyalbadis (H2S) warmoqmnas.

5) samSaqriani rkinis (TSI) metabolizms.

Bbrucelas saxeobebis mgrZnobeloba saRebavebis mimarT:

amisaTvis cdebis proceSi Cvens mier gamoyenebuli iqna fuZe

fuqsini da Tionini.

baqteriologiuri kvleva: brucelas kulturis gamoyofa

iTvleba brucelozis diagnostirebis `oqros standartad~.

brucelebis gamoyofa da identifikacias vaxdendiT

baqteriologiuri gamokvlevis Semdegi meTodebiT:

xelovnur sakveb areebze zrda: brucelebis gamosayofad cdebSi

gamoviyeneT xelovnuri sakvebi niadagebi: maT Soris amJamad

mowodebuli da danergili sakvebi areebi:

a) 5%-iani sisxliani agari

b) brucelas seleqtiuri (fareli) agari brucelas danamatiT.

brucelas seleqtiuri (fareli) (g/l)

xaris gulis infuzia ------------500 g.

triptoza ----------10.g

sodiumis qloridi ------------5.g

baqteriologiuri agari ---------15.g

Jelatini -------------1.g

42

glukoza ------------2.5g

OXOID-is brucelas seleqtiuri danamati SR 0083 formula:

nistatini ---------5000 erTeuli

bacitracini ---------12500 erTeuli

polimiqsini B---------2500 erTeuli

cikloheqsimidi---------10 mg.

nalidiqsis mJava-------2.5 mg.

mikroskopuli gamokvleva: moicavda Cvens mier gamoyofili

brucelas kulturidan damzadebuli nacxis SeRebvas gramis da

kozlovskis meTodiT.

fagoidentifikacia: B. abortus-is tipirebisaTvis Cvens mier

gamoyenebuli iyo Tb fagi.

B. abortus-is kulturis gamozrdas vaxdendiT, anaerostatSi 10-15%

CO2-is Semcvelobis pirobebSi B. abortus mikroaerofiluri

Tvisebis gaTvaliswinebiT.

SratSi aglutinaciisa da akriflavinis testi: brucelas kul-

turis xorklianobis da sigluvis gansazRvrisaTvis, gamoviyeneT

aglutinaciisa akriflavinis testebi.

serologiuri kvleva: kvlevis proceSi brucelozze sisxlis

Sratebi gamovikvlieT roz-bengal testiT, dadebiTi da saeWvo

Sedegebis dadastureba movaxdineT konkurentuli

imunofermentuli analiziT.

43

rZis rgoluri reaqcia: furebis rZis nimuSebi gamovikvlieT

rZis rgoluri reaqciiT.

Ggenetikuri kvleva: polimerazuri jaWvuri reaqciiT

gamovikvlieT B. abortus kulturebi da sisxlis Sratebi.

cdebis procsSi gamoyenebuli diagnostikumebi:

1)GSratebis roz-bengal analizisaTvis diagnostikumad

viyenebdiT, veterinaruli daniSnulebis, ukrainuli warmoebis,

brucelozur ferad, komerciul antigens roz-bengal

testisaTvis. s-#1, k-#1 07.11.11w.

2) konkurentuli imunofermentuli analizisaTvis cdebis

dadgmis procesSi gamoviyeneT, veterinaruli daniSnulebis,

Sveduri warmoebis, svanovas firmis diagnostikumi, P06206,

04.09.11w.

3) furebis rZis nimuSebis gamokvleva xdeboda rZis rgoluri

reaqciiT. diagnostikumad gamoyenebul iqna ukrainuli

warmoebis, brucelozuri antigeni, romelic gankuTvnilia

veterinaruli miznebisaTvis. s-#2, k-#2 23.12.11.w

4) cdebis proceSi polimerazuli jaWvuri reaqciiT brucelas

deteqciisaTvis gamoyenebul iqna, amerikuli warmoebis, qaijenis

mini kiti. (QIA gen Mini kit). lot # 496811, 25.05.2012w.

44

3.2. saqarTveloSi 2005-2011w.w. msxvilfexa pirutyvis

bruceloziT daavadebis dinamika

msxvilfexa pirutyvis brucelozi gavrcelebulia

saqarTvelos mTels teritoriaze metnaklebi intensiobiT. ufro

metadaa dainficirebuli aRmosavleT saqarTvelos

mecxoveleobis fermebisa da mosaxleobis kerZo mflobelobaSi

myofi pirutyvis suladoba. dasavleT saqarTveloSi es infeqcia

ver poulobs masiur gavrcelebas, vinaidan mecxoveleoba

ZiriTadad damxmare dargad aris warmodgenili, dabalia

cxovelTa koncentracia, rac amcirebs maTi

urTierTdainficirebis risks.

aRmosavleT saqarTveloSi epizootiuri situaciis sirTules

sxva mizezebTan erTad mniSvnelovan wilad ganapirobebs

cxovelTa momTabareoba sezonur saZovrebze, gadasarek

trasebze, dasawyurvebel adgilebSi, uSualod saZovrebze,

romlebic ar aris mkacrad izolirebuli erTmaneTisagan.

keTilmouwyobel cxovelTa sazafxulo sadgomebSi xSiria

kontaqti brucelozze arakeTilsaimedo cxovelebis agreTve

gaurkveveli epizootiuri situaciis mqone jgufis cxovelebisa.

saqarTveloSi dRes-dReobiT dabal doneze rCeba brucelozis

sawinaaRmdego sistematiuri, mizanmimarTuli da qmediTi

RonisZiebebis gatareba. droulad ar xdeba daavadebuli

cxovelebis gamovlineba da izolireba, rac warmoSobs

brucelozis gavrcelebis saSiSroebas cxovelebSi da

adamianebSi.

45

am mizniT gavaanalizeT 2005-2011w.w. statistikuri masala da

gamokvlevebs dauqvemdebareT saqarTvelos regionebSi

mosaxleobis kerZo sakuTrebaSi arsebuli msxvilfexa pirutyvis

375426 sisxlis Srati da 2191 furis rZe.

maT Soris aRmosavleT saqarTveloSi (maT Soris Tbilisi),

275599 msxvilfexa pirutyvis sisxlis Srati. dasavleT

saqarTveloSi 99827 sisxlis Srati.

statistikuri masalis gaanalizebam gviCvena (cxrili 3)

saqarTvelos regionebSi (maT Soris TbilisSic), roz-bengal

sinjiT 2005 wels gamovlinda 353 daavadebuli msxvilfexa

pirutyvi, rac Seadgens 0,67%-s, 2006 wels 427 (1,08%), 2007 wels

2054 (3,61%), 2008 wels 1271 (3,61%), 2009 wels 2474 (4,82%), 2010

wels 2247 (1,95%), 2011 wels 667 (3,78%).

46

cxrili 3.

saqarTveloSi 2005-2011w.w. msxvilfexa pirutyvis

bruceloziT daavadebis dinamika

wlebi gamokvleuli m.r.p-

is suladoba

janmrTeli cxovelebis suladoba

daavadebuli cxovelebis suladoba

daavadebuli cxovelebi (%)

2005 53660 53307 353 0.67

2006 39852 39425 427 1.08

2007 56943 54889 2054 3.61

2008 40179 38908 1271 3.17

2009 51417 48943 2474 4.82

2010 115703 113456 2247 1.95

2011 17672 17005 667 3.78

sul 375426 355897 9493 2,53%

47

diagrama 1

saqarTveloSi 2005-2011w.w. msxvilfexa pirutyvis bruceloziT

daavadebis dinamika

msxvilfexa pirutyvis bruceloziT daavadebis SedarebiT maRali

intensivoba aRiniSna 2007, 2008, 2009 da 2010 wlebSi. (diagrama 1).

2005 weli15%

2006 weli11%

2007 weli 15%

2008 weli11%

2009 weli14%

2010 weli32%

2011 weli2%

48

2005-2011w.w.-s monacemebiT regionebis mixedviT brucelozis

gavrcelebis maCvenebeli Semdegi saxiT aris warmodgenili.

(diagrama 2.)

kaxeTis regionSi 2005-2011w.w.-Si roz-bengal sinjiT

gamokvleuli iqna msxvilfexa pirutyvis 48447 sisxlis Srati, maT

Soris dadebiTi iyo-2127.

amave periodSi qvemo qarTlSi gamokvleul iqna 104919

msxvilfexa pirutyvis sisxlis Srati dadebiTi aRmoCnda _1771.

Sida qarTlis regionSi gamokvleuli 13992 Sratis nimuSidan,

dadebiT iyo – 230.

mcxeTa-mTianeTSi gamokvleuli 16258 msxvilfexa pirutyvis

Sratidan dadebiTi aRmoCnda 603.

samcxe-javaxeTSi msxvilfexa pirutyvis 63023 nimuSis

gamokvlevisas dadebiT iyo 747 sisxli Srati.

imereTis regionSi 2005-2011w.w-Si 52922 msxvilfexa pirutyvis

Sratis roz-bengal sinjiT gamokvlevis Sedegad miRebul iqna

1157 dadebiTi nimuSi.

rac Seexeba dasavleT saqarTvelos danarCen regionebs

msxvilfexa pirutyvis sisxlis Sratebis roz-bengal sinjiT

gamokvlevis Sedegad dadebiTi Sedegebi SedarebiT mcire

aRmoCnda (gamonaklisia imereTi) vidre aRmosavleT saqarTvelos

regionebSi, kerZod: guriis regionSi gamokvleuli 10462 Sratis

nimuSidan dadebiTi aRmoCnda – 67.

49

diagrama 2

regionebis mixedviT 2005-2011ww brucelozze mrp-is gamokvlevis maCveneblebi

48447

104919 1399216258

6302352922

10462 11132 4394 20918

28960

2127

1771 230603

7471157

67 72 7 192

2720

sul dadebiTi

50

statistikuri masalis gaanalizebiT procentulad, (cxr. 4) Cans

rom, 2005-2011 wlebSi roz-bengal sinjiT msxvilfexa pirutyvis

bruceloziT daavadebis yvelaze meti SemTxvevaa gamovlenili

TbilisSi-9,4%, regionebidan roz-bengal sinjiT msxvilfexa

pirutyvis bruceloziT daavadebis yvelaze meti SemTxveva

fiqsirebulia mcxeTa-mTianeTSi-3,71%, kaxeTSi-4,39%, imereTSi-2,19%,

Sida qarTlSi-1,69%, qvemo-qarTlSi-1,65, samcxe-javaxeTSi-1.19%.

daaxlovebiT erTnairi Sedegebia danarCen regionebSi: aWara-0,92%,

samegrelo-z. svaneTi-0,65%, guria-0,64%, raWa-0,16%.

Sedegebis analiziT irkveva rom, msxvilfexa pirutyvis roz-

bengal sinjiT dadebiTi SemTxvevebis umetesoba procentulad

modis Tbilissa da aRmosavleT saqarTveloze. dasavleT

saqarTvelodan gamonaklisia imereTi, sadac daavadebis SedarebiT

maRali procentuli SemTxvevaa fiqsirebuli-2,19%.

samegrelo-zemo svaneTSi 11132 gamokvleuli nimuSidan miRebul

iqna 72 dadebiTi sisxlis Srati.

raWaSi gamokvleuli msxvilfexa pirutyvis 4393 Sratis

nimuSidan dadebiTi iyo _ 7.

aWaraSi gamokvleuli 20918 nimuSidan dadebiTi aRmoCnda 192.

TbilisSi gamokvleul iqna msxvilfexa pirutyvis 28960 Sratis

nimuSi dadebiTi iyo – 2720.

2005-2011w.w. gaanlizebuli masalidan aRmosavleT saqarTveloSi

brucelozis SemTxvevebi Warbobs (2,09%), dasavleT saqarTveloSi

brucelozis SemTxvevaTa ricxvs (1,5%).

51

# regioni

2005 2006 2007 2008 2009 2010 2011 %

sisxli

dadebiTi

sisxli

dadebiTi

sisxli

adadebiTi

sisxli

dadebiTi

sisxli

dadebiTi

sisxli

dadebiTi

sisxli

dadebiTi

1 kaxeTi 3542 125 1019 111 20100 746 6710 325 1704 458 14371 261 1001 101 4.39

2 qv. qarTli 15093 63 991 140 4658 64 6074 316 16638 332 55800 566 5665 90 1,69

3 Sida qarTli 10 _ 46 8 1973 30 4443 55 2888 19 2942 64 1690 54 1,65

4 mcxeTa-mTianeTi

6379 124 2781 68 1067 36 220 22 1254 73 3614 251 943 29 3,71

5 samcxe-javaxeTi

_ _ 31449 33 9581 45 5887 18 8902 497 6072 117 1132 37 1,19

6 imereTi 17526 40 1406 35 1867 43 4213 174 8039 324 18391 455 1480 86 2,19

7 guria _ _ 42 0 2056 10 3224 3 1824 26 2255 19 1061 9 0,64

8 samegreloz.svaneTi

_ _ _ _ 1901 2 3692 3 2624 21 1925 36 990 10 0,65

9 raWa _ _ 200 0 269 0 1806 0 401 0 745 0 972 7 0,16

10 aWara 11110 1 70 10 1534 30 828 7 403 0 5934 127 1039 17 0,92

11 Tbilisi _ _ 1848 22 11937 1048 3082 348 6740 724 3654 351 1699 227 9,4

sul 53660 353 39852 427 56943 2054 40179 1271 51417 2474 115703 2247 17672 667

saqarTveloSi mrp bruceloziT daavadebis dinamika regionebis mixedviT cxrili 4

(2005-2011w.w.)

52

2005-2010w.w. furebis rZis rgoluri reaqciiT gamokvlevis Sedegad

miRebulia Semdegi Sedegebi (cxrili 5). 2005 wels dadebiTi

SemTxveva ar dafiqsirebula, (gamokvleuli 30 furebis rZidan),

2006 wels dadebiTi aRmoCnda 8 rZis sinji, (9,76%), 2007 wels

dafiqsirda 21 dadebiTi SemTxveva, rac Seadgens gamokvleuli

rZis 4,62%-s. 2008 wels dadebiTi aRmoCnda gamokvleuli furebis

rZis 9 nimuSi _ 4,31%. 2009 wels 26 dadebiTi anu 6,57% SemTxveva

dafiqsirda, 2010 wels dadebiTi Sedegi dadginda 21 (2,79%)

SemTxvevaSi, 2011 wels dadebiTi iyo 18 sinji, (6,77%). saerTo

jamSi gamokvleuli 2191 furebis rZidan rgoluri reaqciiT

dadebiTi aRmoCnda 103 msxvilfexa pirutyvi, ramac Seadgina 4,71%.

statistikuri masalis analiziT saqarTveloSi roz-bengal

sinjiT da konkurentuli imunofermentuli analiziT 375426 suli

msxvilfexa pirtyvidan daavadebuli aRmoCnda 7245 (2,58%), xolo

2191 rZis nimuSis rgoluri reaqciiT gamokvlevis Sedegad 103

(4,71%). aRmosavleT saqarTveloSi gamokvleuli 275599 msxvilfexa

pirutyvidan dadebiTad moreagire aRmoCnda 5750 (2,09%). xolo,

dasavleT saqarTveloSi _ 99827-dan, 1495 (1,5%).

53

cxrili 5

2005-2011w.w. furebis rZis rgoluri reaqciiT

gamokvlevis Sedegebi

weli gamokvleuli rZis

nimuSebis raodenoba

uaryofiTi

dadebiTi %

2005 30 30 _ _

2006 82 74 8 9,76

2007 455 434 21 4,62

2008 209 200 9 4,31

2009 396 370 26 6,57

2010 753 732 21 2,79

2011 266 248 18 6,77

sul 2191 2088 103 4,71%

54

ruka 1

msxvilfexa pirutyvis sisxlis Sratebis roz-bangal sinjiT gamokvlevis

Sedegebi 2008-2010 w.w.

Mmsxvilfexa

pirutyvis

raodenoba

raionebis

mixedviT

55

3.3. msxvilfexa pirutyvis sisxlis Sratebis roz-bengal sinjiT

da konkurentuli imunofermentuli analiziT gamokvlevis

Sedegebi

gamokvlevebi iTvaliswinebda saqarTvelos soflis meurneobis

saministros laboratoriaSi Semosuli msxvilfexa pirutyvis

sisxlis nimuSebidan Sratebis gamoyofas da maT gamokvlevas

roz-bengal sinjiT. dadebiTi da saeWvo Sratebi, moqmedi

intruqciis Sesabamisad eqvemdebareboda damatebiT gamokvlevas

konkurentuli imunofermentuli analiziT.

roz-bengal sinjis dasadgmelad naWdevian minaze an cremlis

formis mqone myar baraTze an aseTive keramikul dafaze

mikropipetis saSualebiT Segvqonda 30 mkl. (0,03 ml) gamosakvlevi

sisxlis Srati. mis gverdiT vaTavsebdiT amave moculobis

specialur roz-bengalis antigens. Cxiris daxmarebiT wveTebs

erTmaneTSi vurevdiT erTgvarovani, homogenuri siTxis misaRebad,

reaqciis Sedegebs aRvricxavdiT 4 wuTis Semdeg vizualurad,

SeuiaraRebeli TvaliT. paralelurad reaqciis Sedegebis

sarwmunoebisaTvis, analogiuri principiT vaxdendiT

sakontrolo – winaswar cnobili brucelozze dadebiTi da

uaryofiTi Sratebis gamokvlevas. Sefasebas vaxdendiT

sakontrolo Sratebis daTvalierebiT. dadebiT SratTan

specifikuri reaqciis, xolo uaryofiT SratTan uaryofiTi

reaqciis miRebis SemTxvevaSi. reaqciis Sefasebisas

vxelmZRvanelobdiT SratSi aglutinatis warmoqmnis mixedviT.

dadebiTi reaqciis dros mimdinareobda antigenis Sewebeba,

56

romelic mkveTrad moCanda vizualurad wiTel fonze TeTri

fifqebis saxiT.

uaryofiTi reaqciis SemTxvevaSi aglutinati ar warmoiqmneboda,

wveTi rCeboda homogenuri (erTgvarovani). (sur. 2).

sur. 2 roz-bengal testi : 1; dadebiTi kontroli, 2; uaryofiTi

kontroli, sakvlevi nimuSebi.

57

Cvens mier gamokvleuli yvela roz-bengal dadebiTi Sratebis

dadastureba moxda konkurentuli imunofermentuli analiziT.

konkurentuli enzimuri imunofermentuli analizi (C-ELISA),

gamoiyeneba Sinauri da gareuli cxovelebidan aRebul sisxlis

SratebSi Brucella abortus da melitensis-is sawinaaRmdego

antisxeulebis aRmosaCenad.

konkurentuli imunofermentuli analizis dasadgmelad

gamoiyeneba fosoebiani mikrotitraciis planSetebi, romelic

amovsebulia Brucella abortus-is antigeniT (gluvi

lipopolisaqaridiT S-LPS). mikropipetis saSualebiT vamatebdiT

50 mkl. brucelozis sawinaaRmdego antisxeulebis Semcvel

sakvlev Srats. sakvlevi Sratis garda planSetze Segvqonda

Sratis kontrolebi (dadebiTi, sustad dadebiTi, uaryofiTi da

koniugat kontroli). Semdeg yvela fosoSi vamatebdiT 50 mkl.

Tagvis monoklonur antisxeulebs (mAb) (monoklonuri

antisxeulebi specifiuria S-LPS antigenis Teopolisaqaridis

epitopis mimarT). planSets 5 wuTis ganmavlobaSi vaTavsebdiT

Semrevze reaqtivebis ukeT Serevis mizniT. sainkubaciod planSets

vtovebdiT oTaxis temperaturaze (+18-250C) 30 wuTis ganmavlobaSi.

inkubirebis damTavrebis Tanave mikroplanSets vrecxavdiT da

vamatebdiT Txis, Tagvis IgG sawinaaRmdego antisxeulebis

koniugats (100 mkl) pirSuSxas peroqsidazasTan erTad, romelic

ukavSirdeba nebismier mAb-s, es ukanaskneli dakavSirebulia

planSetis S-LPS-Tan. planSets ganmeorebiT sainkubaciod

vtovebdiT oTaxis temperaturaze (+18-250C) 30 wuTis ganmavlobaSi.

58

vaxdendiT planSetis 4-jer garecxvas sarecxi buferiT. momdevno

etapze planSetis TiToeul fosoSi vumatebdiT 100 mkl.

substratis xsnars da sainkubaciod vaTavsebdiT 10 wuTi oTaxis

temperaturaze (+18-250C), reaqciis Sevwyvetis mizniT vamatebdiT 50

mkl. SemaCerebel xsnars, reaqcias vkiTxulobdiT kompiuteris

saSualebiT,Ooptikur simkvrives vzomavdiT mikroplanSetis

fotometris 450 nm-ze.

bruceloziT daavadebuli cxovelidan miRebuli Sratis

damatebisas xdeboda antisxeulebis dakavSireba fosoSi amofenil

antigenTan.

SeukavSirebeli masala ganicdida mocilebas garecxviT

substratis xsnaris damatebamde.Fferis warmoqmna (momwvano-

lurji) warmoqmna xdeboda koniugatis mier substratis

konversiis Sedegad. sakvlev SratSi anti-brucela antisxeulebis

ararsebobisas (uaryofiTi), mAb ukavSirdeba S-LSP antigenis O-

polisaqaridis epitops, rac gamovlindeba feris warmoqmniT

(momwvano-lurji). Tu sakvlevi Srati Seicavs brucela-specifiur

antisxeulebs (dadebiTi), isini konkurencias uweven mAb-s

epitopis adgilebisaTvis da ainhibireben mAb-is SekavSirebas S-

LPS antigenis O-poloisaqaridul nawilTan da Sefervis

warmoqmnas. (sur. 3) e.i. dadebiTi reaqciis SemTxvevaSi feri ar

warmoiqmneba.

konkurentuli imunofermentuli analizis universalur

Tvisebad iTvleba is, rom Stami 19-iT vaqcinirebuli Zroxebidan

miRebuli Sratebi ar Sedian mAb-Tan konkurenciaSi, maTi

59

specifiurobis da dabali afinurobis gamo, rac gvaZlevs

uaryofiT reaqcias. zog SemTxvevaSi gamoricxuli ar aris

vaqcinaciidan 6 Tvis ganmavlobaSi aRebulma nimuSebma gamoiwvion

dadebiTi reaqcia.

sur. 3. C-ELISA.

C++ dadebiTi kontroli

C+ sustad dadebiTi kontroli

C- uaryofiTi kontroli

Cc koniugat kontroli.

60

soflis meurneobis saministros laboratoriaSi 2008 wlis

ganmavlobaSi msxvilfexa pirutyvis brucelozze gamosakvlevad

Semosuli 3082 sisxlis Sratis sinjidan, roz-bengal testiT

dadebiTi aRmoCnda 348, anu 11.30%, 2009 wels brucelozze

gamokvleuli msxvilfexa pirutyvis 6740 sisxlis Sratis

sinjidan, roz-bengalis testiT dadebiTi aRmoCnda 724, (10.75%).

xolo 2010 wels msxvilfexa pirutyvis 3654 sisxlis Sratidan,

roz-bengal testiT dadebiTi Sedegebi aRiricxa 351 SemTxvevaSi,

(9.61%). 2011 wels Semosuli da gamokvleuli 1699 sisxlis

Sratidan, roz-bengal testiT dadebiTebma – 227, anu 13.36%

Seadgina. (diagrama 3.)

yvela zemoT CamoTvlili roz-bengal dadebiTi Sratebis


Sedegebi identuria. rac miuTiTebs aRniSnuli meTodebis

sizustesa da specifiurobaze. (cxrili 6).

61

cxrili 6

msxvilfexa pirutyvis sisxlis Sratebis roz-bengal sinjiT da konkurentuli

imunofermentuli analiziT gamokvlevis Sedegebi

weli sisxlis Sratebis, sinjebis raodenoba

r.b.r ELISA-Ab

dad

ebiT

i (%

)

uar

yofiT

i

dad

ebiT

i

uar

yofiT

i

dad

ebiT

i

2008 3082 2734 348 2634 348 11.30%

2009 6740 6016 724 6016 724 10.75%

2010 3654 3303 351 3303 351 9.61%

2011

1699

1472

227

1472

227

13.36%

62

diagrama 3

3082

6740

3654

1699

348724

351 227

2008w 2009w 2010w 2011w

msxvilfexa pirutyvis brucelozze Sratebis roz-bengal sinjiT da C-

ELISA-Ti gamokvlevis Sedegebi

sul dadebiTi

63

3.4. furebis rZis brucelozze rgoluri reaqciiT gamokvlevis

Sedegebi

cdebis msvlelobis periodSi furebis rZis gamokvlevas

vaxdendiT rZis rgoluri reaqciiT, Semdegi wina pirobebis

gaTvaliswinebiT: gamosakvlevad vargisia, mxolod mouxdeli da

axali rZe. aRniSnul reaqciaSi gamokvlevas ar eqvemdebareba rZis

nimuSi, romelic aRebulia cxovelis axurebisa da dagrilebis

periodSi, nayofis mogdebidan pirveli Tormeti dRis

ganmavlobaSi, agreTve mastitiT daavadebuli furebis rZe.

gamokvlevebs davuqvemdebareT saqarTvelos sxvadsxva

raionebidan 2008-2011w.w. soflis meurneobis saministros

laboratoriaSi Semosuli mosaxleobis kerZo sakuTrebaSi

arsebuli 97 furis rZe.

gamokvlevis win rZian WurWels vanjRrevdiT naRebis Tanabari

ganawilebisaTvis, serologiur sinjarebSi Segvqonda 2000 mkl.

(an 2-2 ml.) rZe da vumatebdiT 200 mkl. (an 0,2 ml.) antigens.

sinjarebs guldasmiT vanjRrevdiT.

ZiriTadi reaqciis paralelurad sarwmuno Sedegebis misaRebad

vdgavdiT Semdeg kontrolebs:

_ winaswar cnobili janmrTeli cxovelis rZesTan,

_ janmrTeli cxovelis rZisa da dadebiTi Sratis narevTan.

Stativs gamosakvlevi da sakontrolo rZis sinjarebiT

vaTavsebdiT TermostatSi 370C erTi saaTis ganmavlobaSi.

64

reaqciis Sefasebas vaxdendiT vizualurad, Termostatidan

sinjarebis gamoRebisTanave. Sedegebis aRricxvas viwyebdiT

sakontrolo sinjarebidan. reaqcia janmrTeli cxovelis rZesTan

uaryofiTia, xolo janmrTeli cxovelis rZisa da brucelozis

dadebiTi Sratis narevTan-dadebiTi. reaqciis Sedegebis

Sefasebas vaxdendiT jvrebiT:

+++ (sami jvari) _ rZis svetis zemoTa nawilis naRebis fenaSi

mkafiod gamoxatulia lurji feris rgoli, rZis danarCeni masa

TeTria.

++ (ori jvari) _ naRebis fenaSi naTlad gamoxatulia lurji

rgoli, rZis danarCeni masa molurjo Seferilobisaa.

+ (erTi jvari) _ rZis sveti SeRebilia lurj ferSi, naRebis fena

TeTri da moyviTaloa.

rZis sinji, romelic Sefasda sami an ori jvriT _ dadebiTia,

xolo erTi jvriT _ uaryofiTi. (sur. 4).

brucelozze rZis nimuSebis, rgoluri reaqciiT Catarebulma

gamokvlevebma gviCvena, rom dadebiTi SemTvevebis umetesoba

aRmosavleT saqarTveloze modis. 97 rZis sinjidan 17 SemTxvevaSi

rgoluri reaqcia aRmoCnda dadebiTi, maT Soris 2008w. -1, (16.67%),

2009w. – 4, (33.34%), 2010w. -7, (18.92%), 2011w– 5, (10,81%) (cxr. 7).

(diagrama 4).

aRsaniSnavia, rom Teqvsmetive dadebiT SemTxvevaSi aRiricxa

mkveTrad dadebiT reaqciad (+++), rasac adasturebs sinjaris

zeda nawilSi, naRebis fenaSi lurjad SeRebili damaxasiaTebeli

65

rgolis warmoqmna, rac advilad gasarCevia rZis svetis TeTr

fonze.

a) b) g)

sur. 4. rZis rgoluri reaqcia.

a) dadebiTi kontroli.

b) uaryofiTi kontroli.

g) sakvlevi nimuSi.

66

cxrili 7.

furebis rZis rgoluri reaqciiT gamokvlevis Sedegebi

weli gamokvleuli rZis

nimuSebis raodenoba

uaryofiTi

dadebiTi %

2008 6 5 1 16,67%

2009 12 8 4 33,34%

2010 37 30 7 18,92%

2011 42 37 5 11,9%

sul 97 80 17 17.52%

67

diagrama 4

furebis rZis rgoluri raqciiT gamokvlevis Sedegebi

(2008-2011w.w.)

2008w 2009w 2010w 2011w

6

12

37 42

1

4

7 5

sul dadebiTi

68

3.5. furebidan gamoyofili B. abortus kulturebis Seswavlis

Sedegebi

2010-2011 w.w.-Si Catarebuli kvleviTi samuSaoebi

iTvaliswinebda msxvilfexa pirutyvidan gamoyofili brucelozis

aRmZvrelebis tipirebas. cdebis procesSi gamokvlevas

davuqvemdebareT soflis meurneobis saministros laboratoriaSi,

saqarTvelos sami regionidan (qvemo qarTli, kaxeTi da imereTi)

2008-2010w.w. Semosuli 1253 sisxlis, 577 rZis, 5 mogdebuli nayofis,

da 53 Sratis nimuSi. Brucella-is gamoyofisa da

identifikaciisaTvis, vaxdendiT sisxlis, rZis da mogdebuli

nayofidan aRebuli nimuSebis, CaTesvas xelovnur sakveb areebze,

kerZod brucelas seleqtiur niadagze da sisxlian agarze.

Brucella saeWvo kulturebs vikvlevdiT bioqimiuri testebiT:

oqsidaza, katalaza, ureaza, gogirdwyalbadis warmoqmna da

samSaqriani rkinis metabolizmi. Brucella-as koloniebis

morfologiis dasadgenad gamoviyeneT akriflavinis da kristal-

violetis testebi, movaxdineT miRebuli kulturebis

fagotipireba Tb fagiT. yvela zemoT CamoTvlili testebis

gamoyenebis Sedegad gamoyofil iqna B. abortus 20 saeWvo kultura,

romelTa dadastureba moxda myisieri PCR-iT da AMOS-PCR-iT.

paralelurad movaxdineT 53 Sratis nimuSis eqstragireba,

miRebuli d.n.m-is nimuSebi gamokvleul iqna myisieri PCR-iT da

AMOS-PCR-iT, 31 dadebiTi SedegiT. (algoriTimi #1).

69

algoriTmi 1

d.n.m-is

eqstraqcia

morfologia bioqimia H2S; TSI mgrZnobeloba fagotipireba saRebavebze

paTologiuri masala

(sisxli, rZe, mogdebuli nayofi)

CaTesva sakveb areebze

sisxliani, agari,

seleqtiuri niadagi

(fareli)

Aakriflavini,

kristal violeti

PCR

AMOS PCR

oqsidaza,+ ureaza,+

katalaza+

TB fagi

+

fuZe fuqsini+ Tionini _

mikrosko-

pia

70

3.5.1. morfologia da tinqtorialuri Tvisebebi

B. abortus Stamebis morfologiisa da tinqtorialuri Tvisebebis

dasadgenad paTologiuri masalidan da xelovnur sakveb areebze

(brucelas bulioni, farelis agari) nazardi kulturebidan

davamzadeT nacxebi da SevRebeT gramis da kozlovskis wesiT, es

ukanaskneli tipuria da Sereul masalaSi brucelebis sxva

mikrobebisagan diferencirebis mizniT gamoiyeneba.

grami wesiT SeRebva. Ggramis wesiT SeRbvisaTvis brucelebis 48

saaTiani koloniebidan damzadebuli nacxebi gavaSreT, movaxdineT

fiqsacia gacxelebiT, fiqsirebul nacxebs davafareT filtris

qaRaldi, davasxiT gencian-violetis saRebavi; davayovneT 2 wuTi,

Semdeg movacileT filtris qaRaldi da davawveTeT lugolis

xsnari, davayovneT 2 wuTi; 20 wamis ganmavlobaSi preparati

davamuSaveT 960-iani spirtiT, nacxi CavrecxeT wyliT, SevRebeT

safraniniT 2 wuTis ganmavlobaSi, kvlav CavrecxeT onkanis

wyliT, gavaSreT saSrobi qaRaldiT da gamovikvlieT mikroskopSi

imersiuli sistemiT. Cvens mier gamoyofili da Seswavlili B.

abortus Stamebi, garmuaryofiTi, wiTel ferSi SeRebili

kokobaqteriebia, romlebic preparatSi umetesad dalagebulia

wyvilad da gamonaklisad erTeuli saxiT. (sur. 5)

kozlovskis meTodiT SeRebva. Aam mizniT fiqsirebul nacxebs

vawveTebdiT safraninis 2%-ian wylian xsnars, da vRebavdiT

gacxelebiT spirtquris alze airebis pirveli buStukebis

gamoyofamde. preparati CavrecxeT wyliT. nacxi damatebiT

SevRebeT 0,75-1%-iani meTilenis lurjis wyliani xsnariT 30 wamis

71

ganmavlobaSi. Ppreparatebi ganmeorebiT CavrecxeT wyliT,

gavamSraleT da gavsinjeT mikroskopSi, imersiuli sistemiT.

mikroskopuli suraTi- Cvens mier gamoyofili B. abortus

kokobaqteriebi aRmoCnda SeRebili Ria vardisferSi, rac maTi

tipurobis maCvenebelia.

suraTi 5. gramis wesiT SeRebili brucelebi

72

2.5.2. sakveb areebze zrdis Tavisebureba

myar sakveb areebze zrdis mixedviT arCeven brucelebis S –

tipiur_gluvi, R – Secvlil_xorkliani, da M – lorwovan_

koloniebs. brucelebisaTvis gamonaklisad damaxasiaTebelia

filtrSi gamavali e.w. L formis warmoqmna.

tipiuri virulenturi Stami S-forma, agaris zedapirze

warmoqmnis wvril, 2-3 mm. diametris, mrgval, amoburcul, gluvi

zedapirisa da swori kideebis mqone, mbrwyinav, Taflisfer,

karaqis konsistenciis, koloniebs. brucelas xandazmul

koloniebs aqvs Jangisferi.

avirulenturi (R-forma) agarze warmoqmnis xorklian koloniebs.

msxvilfeaxa pirutyvis savele nimuSebi (anti-koagulantiani

sisxli, rZe, qsovili an mSobiarobasTan asocirebuli masala.)

SesaZlebelia Seicavdes sxva baqteriebsac, Brucalla-s saxeobaTa

sufTa kulturis gamoyofa saWiroebs specilizirebul sakveb

areebs. B. abortus kulturis gamosayofad da zrdis

Taviseburebebis dasadgenad, gamoviyeneT seleqtiuri niadagi,

romelic axdens ucxo mikroorganizmebis zrdis daTrgunvas da

xels uwyobs brcelebis zrda-ganviTarebas. Bbrucella-s seleqtiuri

niadagi Seicavs antibiotikebis narevs. sakvebi are brucelebis

zrdis dasaCqareblad danamatis saxiT Seicavs sisxls, (brucela

–uaryofiTi), an Srats. brucelebis gamosayofad paT. masalis

nimuSebs finjanze gaTesvamde vyinavdiT -200C-ze, gayinva xels

73

uwyobs nebismier baqteriuli ujredebis ganTavisuflebas

sisxlis formiani elementebidan. gayinul nimuSebs valRobdiT

da vorteqsis gamoyenebiT vaxdendiT Serevas.

Txevadi nimuSebi (rogoricaa sisxli, rZe) 100 mkl. raodenobiT

steriluri pipetis wveris saSualebiT gadagvqonda da

vaTavsebdiT seleqtiuri niadagis centrSi.

qsovilis nimuSebs steriluri pincetisa da makratlis

gamoyenebiT vaquvmacebdiT “sufTa kideebis” misaRebad.

steriluri maryuJiT vaxdendiT wveTis an paT. masalis

gadaTesvas finjanze daStrixviT, “molis” msgavsad.

gaSrobis Semdeg finjnebs vaTavsebdiT Tavsaxurian plastmasis

konteinerSi da vtovebdiT TermostatSi 370C-ze. 10-15% CO2-ian

paketebaTan erTad. (B. abortus mikroaerofilia da zrdisaTvis

saWiroebs CO2-iani garemos). finjnebis inkubirebas vaxdendiT 21

dRemde, TermostatSi 370C-ze.

naTesebs vamowmebdiT yovel 24 saaTSi da TiToeul kolonias

vniSnavdiT finjnis fuZeze SemoxazviT, rogorc (ara “brucelas”)

dabinZurebuli kolonia.

Pinkubaciidan me-2-3-e dRes aRiniSneboda mkrTali, gamWvirvale,

Taflisferi Seferilobis 2-3 mm diametris mqone mrgvali

koloniebi, (sur. 6.) isini amoburcul zedapiriani da moTeTroa.

preparatSi qsovilebis nawilakebisa da cximis wveTebis

zemoqmedebis Sedegad aRiniSneboda uswormasworo kideebiani

74

koloniebis zrda. saeWvo koloniebi warmoqmnisTanave gadagvqonda

seleqtiur niadagze Semdgomi gamokvlevebisaTvis.

sur. 6. brucelas koloniebi (S forma)

Semdgomi testirebisaTvis, vaxdendiT saeWvo koloniebis

brucelas seleqtiur niadagze gadaTesvas. (farelis niadagi).

kultivirebisaTvis naTesi kulturebi movaTavseT (CO2-is gareSe)

TermostatSi 370C-ze. zrdis xasiaTze dakvirvebas vaxdendiT

yoveli 24 saaTis Semdeg. gamokvlevebma gviCvena kulturaTa zrda

75

susti nazardis saxiT aRiniSna (sur. 7). mesame dRes da

maqsimalur intensivobas miaRwia 21-e dRes. (cxrili 8).

sur. 7. B. abortus nazardi brucelas seleqtur (farelis) agarze.

76

B. abortus sakveb areebze zrdis maxasiaTelebi cxrili 8

# mikrobis dasaxeleba koloniis warmoqmnis

dro (sT-Si) koloniis forma

koloniis diametri (mm)

1 0178-b 72 S 3 2 051-m 48 S 2 3 285-b 72 S 3 4 N1-t 48 S 2 5 IM.0014-m 48 S 2 6 IM.0018-m 48 S 2 7 K.K.0519-m 72 S 3 8 T.01-m 72 S 3 9 M.T.02-m 48 S 2 10 K.K.0790-m 72 S 3 11 K.K.1102-b 48 S 2 12 K.1556-m 72 S 3 13 K.1601-m 72 S 3 14 K.1733-m 48 S 2 15 K.1745-m 48 S 2 16 K.1573-m 72 S 3 17 IM.0030-m 72 S 3 18 T.03-m 72 S 3 19 K.K.1109-m 72 S 3 20 K.K.0808-m 72 S 3

77

3.5.3 B. Abotus izolatebis bioqimiuri TvisebebiB

B. abortus saxeobis identifikaciisaTvis gamoviyeneT Semdegi

bioqimiuri testebi: oqsidaza, ureaza katalaza; gogirdwyalbadis

gamomuSaveba da rkinis metabolizmi. (cxrili 9)

oqsidazas testi. oqsidazas testiT xdeba ferment citoqrom

oqsidazas aRmoCena, is gamoiyeneba gram-uaryofiTi baqteriebis

bioqimiuri testirebisaTvis. dadebiTi Sedegebisas citoqrom

oqsidazas arsebobis SemTxvevaSi misi reaqtivebi urTierTqmedeben

indofenolis lurjTan.

oqsidazas testis Casatareblad steriluri tamponiT viRebdiT

2-3 izolirebul kolonias. uSualod tamponze arsebul

baqteriaze vawveTebdiT oqsidazas reaqtivs da 30 wamis Semdeg

aRvricxavdiT Sedegebs. Cvens mier gamoyofili B. abortus yvela

izolati oqsidaza dadebiTia, rasac adasturebs aRniSnuli

testis Catarebisas lurji feris warmoqmna.

ureazas testi _ gamoiyeneba mikrorganizmis mier ferment ureazas

saSualebiT Sardovanas hidrolizis TvisebaSi. amoniumis ori

erTeuli warmoiqmneba tute Tvisobriobis Sedegad. tutis

aRmoCena xdeba pH-indikatoris saSualebiT. qristensenis

Sardovana romelic pH indikators fenol wiTels Seicavs,

Cveulebriv mJave garemoSi (pH 6.8) yviTeli Seferilobisaa; tute

garemoSi (pH 8.4) indikatori icvlis fers da xdeba movardisfro-

mowiTalo.

ureazas testis Casatareblad viRebdiT sinjaras qristensenis

78

iribi agariT. agaris zedapirze vaxdendiT brucelas koloniebis

gadaTesvas. sinjarebs inkubirebisaTvis vtovebdiT TermostatSi

35-370C-ze. nazardis Semowmebas vaxdendiT 24-48 saaTis Semdeg.

Catarebuli gamokvlevebiT davadgineT, rom B. abortus yvela Stami,

iwvevs vardisfer-wiTeli feris warmoqmnas, rac dadebiTi ureazas

testis maCvenebelia. (suraTi 8). aRsaniSnavia, rom Cvens mier

gamoyofil StamebSi vardisfer-wiTeli feris warmoqmna

sxvadasxva intensivobiT xdeboda.

sur 8. ureaza testi

79

katalazas testi: fermenti katlaza axdens wyalbadis zeJangis

hidrolizis katalizebas, ris Sedegad warmoiqmneba wyali da

Jangbadi. Cvens mier gamoyofili brucelaTa yvela saxeoba

aRmoCnda katalaza dadebiTi.

katalazas testis Casatareblad steriluri maryuJiT

izolirebuli kolonia gadagvqonda sasagne minaze, romelSic

Segvqonda 1-2 wveTi 3%-iani wyalbadis zeJangis xsnari, rasac

myisierad Tan sdevda airebis buStukebis warmoqmna, rac Cvens

mier gamoyofili B. abortus katalazuri aqtivobis mimaniSnebelia.

samSaqriani rkinis agari (Triple sugar Iron – TSI) misi saSualebiT

xdeba deqstrozas, laqtozasa da saqarozas fermentaciis

dadgena. misi gamoyeneba SesaZebelia damatebiTi testiT zogierTi

mikroorganizmebis mier wyalbadis zeJangis produqciis

aRmosaCenad, Tumca araefeqturia brucelas saxeobebis

dasadgenad. agaris feri ganicdis cvlilebas – yviTeli mJavebis

producirebisas, da wiTeli tutes warmoqmnisas – fermentaciis

dros SesaZlebelia agreTve warmoqmnas gazebi. brucelas

saxeobebi, arian ara-reaqtiuli, anu TSI uaryofiTi

mikroorganizmebi. (ar icvleba Seferiloba da ar xdeba airebis

warmoqmna).

brucelas saxeobaTa identifikaciisaTvis sinjarebSi

samSaqriani rkinis iribi agariT vaxdendiT B. abortus kulturaTa

gaTesvas gazonis saxiT. sacobsa da sinjaris Sida zedapirs

Soris vaTavsebdiT tyviis acetatian strips. sinjarebs vtovebdiT

35-370C-ze 24-48 saaTis ganmavlobaSi. Sedegebze vmsjelobdiT

80

sakvebi aris feris SecvliT, airebis warmoqmniT, rac Cven cdebSi

ar aRmoCnda damaxasiaTebeli B. abortus kulturebisaTvis anu

brucelebis aRniSnuli saxeobebi TSI uaryofiTia.

H2S-is testi: tyviis acetatis test stripebi gamoiyeneba baqteri-

ebis mier gogirdwyalbadis gamoyofis aRmosaCenad.

gogirdwyalbadis warmoqmna SesaZlebelia brucelas mier

gogirdis Semcveli aminomJavebis daSlis Sedegad. amisaTvis

baqteriaTa kulturebi CavTeseT karborhidratebian sakveb

niadagze, kerZod samvalentiani Saqris rkina. gogirdwyalbadi

Sevida ra kontaqtSi tyviis acetatis stripTan, warmoqmna Savi

precipitati, rac stripze Savi feris reaqciis warmoqmniT

gamoixata. B. abortus gogirdwyalbad dadebiTia. (suraTi. 9)

sur. 9. gogorgwyalbadis testi.

81

Sratis aglutinaciis testi: Sratis aglutinaciis testi

gamoiyeneba mikrobebSi O-polisaqaridis gverdiTi jaWvis

arsebobis dasadgenad, rac damyarebulia homogenuri

antisxeulebis SeboWvaze. O-polisaqaridis gverdiTi jaWvis

arsebobis SemTxvevaSi antisxeulisa da antigenis

urTierTqmedebas Tan sdevs aglutinacia. Ggluvi koloniebi

aglutinirdeba O-polisaqarid specifiur poliklonur SratTan,

maSin roca xorkliani mikroorganizmebi darCeba suspenziis

saxiT. B. abortus gluvi mikroorganizmia, romelic aglutinirdeba

Sratis arsebobisas.

Sratis aglutinaciis testisaTvis minaze vawveTebdiT 30 mkl.

Srats, SratSi steriluri maryuJiT Segvqonda 2-3 B. abortus-ze

saeWvo kolonia. ingredientebs guldasmiT urevdiT homogenuli

xsnaris misaRebad. dadebiTi reaqcia miuTiTebs, rom datanili

saeWvo kolonia miekuTvneba B. abortus, rac Cvens cdebSi

damaxasiaTebelia B. abortus yvela izolatisaTvis.

akriflavinis testi: akriflavinis testi gamoiyeneba

mikroorganizmTa gluvi an xorkliani koloniebis

gansazRvrisaTvis. xorkliani koloniebi ganicdian aglutinacias

neitraluri akriflavinis arsebobis SemTxvevaSi. am movlenas

safuZvlad udevs reaqciaSi maTi gareTa membranis sruli CarTva.

xorkliani mikroorganizmebi ganicdian aglutinacias

akriflavinis arsebobisas; mis sapirispirod gluvi

mikroorganizmebi aRniSnul Tvisebas moklebulia.

akriflavinis testis Casatareblad sufTa sasgne minaze Segv-

82

qonda akriflavinis xsnari 30 mkl-is moculobiT, steriluri

maryuJiT wveTSi vxsnidiT 2-3 B. abortus-ze saeWvo kolonias,

homogenuri xsnaris misaRebad. komponentebs erTmaneTSi

guldasmiT urevdiT. B. abortus, rolebic warmoqmnian

gluvzedapirian kolonias ar ganicdis aglutinacias da xsnarSi

imyofebian fifqebis saxiT.

fuZe fuqsinisa da Tioninis testi: gamoiyeneba brucelas

saxeobaTa identifikaciisaTvis. fuZe fuqsinis Semcvel sakveb

areebSi B. abortus kulturebi ganicdian intensiur zrdas; Tioninis

Semcvel sakveb areebSi maTi zrda iTrguneba.

brucelaTa zrda-ganviTarebaze fuZe-fuqsinisa da Tioninis

gavlenis dasadgenad B. abortus kulturebi gavTeseT petris

finjnebSi Camosxmul, aRniSnul nivTierebaTaA Semcvel sakveb

areebze. naTesebi inkubaciisaTvis movaTavseT TermostatSi 370C-

ze. Sedegebi aRvricxeT inkubirebidan 48-72 saaTis Semdeg.

gamokvlevebiT davadgineT, rom Cvens mier paT. masalidan

gamoyofili B. abortus ocive Stami intensiurad izrdeba fuZe

fuqsinis Semcvel sakveb areze, rac Seexeba Tioninian niadagze

zrda ar aRiniSneba. (sur. 10.)

83

a) b)

sur. 10. mgrZnobeloba saRebavze

a) fuZe fuqsini (vardisferi): B. abortus zrda.

b) Tionini (lurji): B. abortus zrda ar aris.

84

B. abortus izolatebis bioqimiuri Tvisebebi cxrili 9

# brucelas kulturebi

gramiT SeRebva

gluvi an xorkliani

CO2 oqsidaza katalaza ureaza H2S TSI Tionini fuZe

fuqsini

1 0178-b uaryofiTi gluvi + + + + + _ _ +

2 051-m uaryofiTi gluvi + + + + + _ _ +

3 285-b uaryofiTi gluvi + + + + + _ _ +

4 N1-t uaryofiTi gluvi + + + + + _ _ +

5 IM.0014-m uaryofiTi gluvi + + + + + _ _ +


7 K.K.0519-m uaryofiTi gluvi + + + + + _ _ +

8 T.01-m uaryofiTi gluvi + + + + + _ _ +

9 M.T.02-m uaryofiTi gluvi + + + + + _ _ +


11 K.K.1102-b uaryofiTi gluvi + + + + + _ _ +

12 K.1556-m uaryofiTi gluvi + + + + + _ _ +






18 T.03-m uaryofiTi gluvi + + + + + _ _ +



85

3.5.4. Br. Abortus bovis izolatebis fagoidentifikacia

brucelaTa fagoidentifikacia specifikuri indikatori

fagebiT xorcieldeba.

brucelaTa specifikuri fagebis miRebis, Seswavlis

medicinasa da veterinariaSi warmatebiT gamoyenebis adreuli

gamokvlevebi ekuTvis m. d. droJevkinas (1953). mkvlevaris mier

miRebuli brucelebis polivalenturi baqteriofagi.

brucelozis aRmZvrelis 174 Stamze SemowmebiT fagi aRmoCnda

specifikuri. baqteriofagi mkvlevarma warmatebiT gamoiyena

brucelebis inaglutinobeluri Stamebis identifikaciisaTvis.

Teoriuli da praqtikuli TvalsazrisiT sagulisxmo cdebia

Catarebuli brucelozis sadiagnostikod fagis zrdis reaqciis

gamoyenebaze. n. n. ostrovskaiam, d. m. goldfarbma (1959) reaqciaSi

indikatorul fagad gamoiyenes B. abortus-is tipospecifikuri fagi.

maT daadgines, rom 72 saaTiani inkubaciis pirobebSi reaqcia

xelovnurad dainficirebul bulionSi, erTeuli mikrobis

aRmoCenis saSualebas iZleva. ftzr SedarebiT naklebad

mgrZnobiarea rZisa da niadagis gamokvlevisas, razedac gavlenas

axdens vulgaruli mikroflora.

sakvebisa da soflis meurneobis organizaciis (FAO).

eqspertebis daskvniT (1986), sxva kompleqsur meTodebTan erTad

brucelas Stamebis diferenciaciisaTvis rekomendebulia

Tbilisis eliavas saxelobis institutSi miRebuli Tb fagis

gamoyeneba. (m. fofxaZe Tanaavt. 1968w).

86

fagotipireba Brucella-is Stamebis identifikaciis damatebiTi

testia Brucella–is fagi da misi rutinuli test-ganzaveba (RTD)

axdens B. abortus-is gluvi koloniebis liziss da iZleva mcire

zomis folaqebs. sixlidan, Sratidan, rZidan da mogdebuli

nayofidan gamoyofili B.abortus kulturebis

fagoidentifikaciisaTvis petris finjanSi farelis niadagis

zedapirze baqteriologiuri maryuJiT vavlebdiT 24-saaTiani

brucelas bulioniani kulturis zonrebs, oTaxis temperaturaze

gaSrobis Semdeg zonrebs wvrilwveriani pipetiT vawveTebdiT Tb

fagis rutinul test-ganzavebis 1 wveTs (25 mkl.) wveTis gaSrobis

Semdeg gadmobrunebul finjnebs sainkubacioT vaTavsebdiT 10%

CO2-is pirobebSi TermostatSi 370C-ze, 48 saaTi.

Catarebulma gamokvlevebma (cxrili 10). gviCvena, rom

Seswavlili yvela oci Stami ganicdis liziss Tb specifikuri

fagiT rac B. abortus arsebobis damadasturebelia. fagiT

kulturaTa lizisis xarisxma 3+ da 4+ Seadgina. (sur. 11)

Seswavlili 20 kulturidan yvelam ganicada Tb fagiT sruli

lizisi, romlis xarisxic Sefasda 4+.

B

87

cxrili 10

B. abortus kulturebis

fagomgrZnobeloba

# kuturaTa dasaxeleba Sedegebi

1 0178-b 4+

2 051-m 4+

3 285-b 4+

4 N1-t 4+

5 IM.0014-m 4+

6 IM.0018-m 4+

7 K.K.0519-m 4+

8 T.01-m 4+

9 M.T.02-m 4+

10 K.K.0790-m 4+

11 K.K.1102-b 4+

12 K.1556-m 4+

13 K.1601-m 4+

14 K.1733-m 4+

15 K.1745-m 4+

16 K.1573-m 4+

17 IM.0030-m 4+

18 T.03-m 4+

19 K.K.1109-m 4+

20 K.K.0808-m 4+

88

sur. 11. B. abortus kulturis Tb fagiT lizisi.

89

3.5.5. B. abortus bovis izolatebis d.n.m skriningi

polimerazuli jaWvuri reaqcia: polimerazuli jaWvuri reaqciis

principi orjaWviani d.n.m-is denaturaciisas erTjaWviani d.n.m-is

miRebaSi mdgomareobs, es procesi mimdinareobs fermentebis

meSveobiT, d.n.m-is mokle komplementarul monakveTs (praimeri)

SeuZlia daukavSirdes d.n.m-is calke myof jaWvis standartul

fragments (matrica). Ppraimeri scnobs matricas da ukavSirdeba

(axdens hibridizacias) amocnobil Tanamimdevrobas. Ppraimeri 3’-

daboloebis gamoyenebiT d.n.m polimeraza axdens d.n.m-is axali

jaWvis sinTezs (elongacia) _ dagrZeleba. d.n.m-is raodenobis

gaormageba 1 ciklis ganmavlobaSi xdeba. 25 ciklis Semdeg 3,2X107

d.n.m-is molekulaa amplificirebuli.

praimerebad gamoiyeneba sinTezirebuli erTjaWviani dnm (20-30

nukleotidi). ori sxvadasxva praimeris seqvensi (gaSifruli

nukleotidebis Tanamimdevroba) gamoiyeneba gasamravlebeli

regionis orive mxridan SemosazRvrisaTvis. erTi praimeri samizne

regionis dasawyisSi komplementuria dnm erTi jaWvisa, meore

praimeri samizne regionis boloSi komplementuria meore jaWvis.

kvlevis procesSi Cvens mier gamoyenebul iqna myisieri

polimerazuli jaWvuri reaqcia, romelic tradiciulis

modifikaciaa, ufro swrafia da gamosayeneblad mosaxerxebeli.

Tumca tests sWirdeba eqstraqciis Catareba, d.n.m-is gamoyofa.

polimerazuli jaWvuri reaqciis msvlelobas vaxorcielebdiT

ramdenime etapad: Tavdapirvelad vaxdendiT nimuSidan (Srati, B.

abortus kultura da a.S) d.n.m-is eqstraqcias.

90

1) d.n.m-is eqstaqcia: 20 mkl. K proteinazas vasxavdiT 1.5 ml-ian

ependorfis sinjaraSi. vamatebdiT 200 mkl. antikoagulirebul

sisxls an 10%-ian qsovilis homogenats. vamatebdiT 200mkl AL

bufers. vorteqsis saSualebiT vaxdendiT maT Serevas da

vaCerebdiT sainkubaciod 560C-ze 10 wuTis ganmavlobaSi.

vamatebdiT 200 mkl. 96-100-ian spirts, isev vavorteqsebdiT. 620

mkl. narevi gadagvqonda Dneasy mini svetSi, vapipetirebdiT da

vacentrifugirebdiT 1wT 6000 x g, sinjaris qveda nawils

vagdebdiT, xolo sinjaris sveti gadagvqonda axal Sesagrovebel

sinjaraSi. Semdeg etapze vamatebdiT 500mkl. AW1 buferi (AW1-es

aris sarecxi buferi, romelic nimuSidan gamorecxavs yvela

zedmet balasts, rogoricaa marilebi an cilovani nivTierebebi).

isev vacentrifugirebdiT 1wT. 6000 x g, sinjaris qveda nawils isev

vagdebdiT, minis svets vdebdiT axal Sesagrovebel sinjaraSi.

vamatebdT 500mkl. AW2 bufers (esec sarecxi buferia, igive

funqcias asrulebs rasac AW1 da gamorecxavs darCenil balasts).

gavimeoreT centrifugireba 3wT. 16,000 x g, sinjaris qveda nawili

gadavagdeT, miRebuli eqstraqtiani minis sveti gadavitaneT

Sesagrovebel sinjaraSi. gavimeoreT centrifugirebis procedura,

mSralad. (16,000 kerZod x g,). miRebili eqstaqtiani minis sveti

gadavitaneT ependorfis sinjaraSi (romelic Tavisufalia r.n.m-

azasagan) da davamateT 200mkl. AE buferi (SeiZleba gamoyenebuli

iqnas r.n.m-azasagan Tavisufali wyali.) gavaCereT 1 wT. oTaxis

temperaturaze da davacentrifugeT 1 wT. 6000 x g. miRebuli

eqtragirebuli d.n.m mzadaa Semdgomi testirebisaTvis (im

SemTxvevaSi Tu PCR-is gaSveba ar xerxdeba maSinve, maSin eqsragi-

91

girebuli d.n.m inaxeba macivarSi -20 an -800 C-ze.)

2) PCR master miqsis momzadeba: brucelas myisieri PCR-iT sadiag-

nostikod gamoiyeneba aidahos teqnologiT damzadebuli kiti,

romelic dafuZnebulia mSral teqnologiaze

`liofilizirebuli~. es imas niSnavs, rom mwarmoeblis mier

damzadebuli master miqsi mowodebulia mSrali saxiT. (master

miqsi Seicavs polimerazas, oligonukleotidebis mokle

praimerebs, oTx dezoqsiribonukleotidebisagan Semdgar

asaSenebel blokebs dnm-sTvis da kofaqtors MgCl2.-s).

liofilizirebuli anu mSrali master miqsis aRdgenas

vaxdendiT 40 mkl. deionizirebuli wylis damatebiT (dadebiTi da

uaryofiTi kontrolebisaTvis).

3) PCR reaqcia: sinjarebSi, ucnobi nimuSebisTvis, vamatebdiT 40

mkl. winaswar, Cvens mier eqtragirebul da ganzavebul sakvlev

d.n.m-s (4 mkl. d.n.m + 36 mkl. deionizirebuli wyali).

4) PCR ciklireba: `laiT cikleris~ kapilarul sinjarebSi

Segvqonda 18 mkl. aRdgenili reagentebi da sakvlevi nimuSebi.

Roche LightCycler 2.0. kapilarTa rotoris maqsimaluri tevadobaa 32

kapilari. Tu dublirebuli dadebiTi da uaryofiTi

kontrolebisaTvis gamoiyeneba oTxi kapilari, SesaZlebelia

dublirebulad gaSvebul iqnas 14 sakvlevi nimuSi. Semdeg es

sinjarebi CavtvirTeT `laiT cikleris~ karuselSi

centrifugirebisaTvis. Semdeg xdeba reaqciis gaSveba. erTi

gaormagebis cikli mimdinareobs 3 etapad:

92

1) denaturacia, 2) hibridizacia, 3) elongacia. samive etapi damo-

kidebulia garkveul temperaturaze.

1)Ddenaturaciis dros dezoqsiribonukleinis mJava cxeldeba

maRal temperaturaze, (94 -970C-ze 1 wuTis ganmavlobaSi). rac

ganapirobebs dezoqsiribonukleinis mJavas ori Zafis

erTmaneTisagan daSorebas.

2) hibridizaciis dros xdeba polimerazuli jaWvuri reaqciis

temperaturis Semcireba 50-650-mde 1 wuTis ganmavlobaSi ise, rom

praimerebs SesaZlebloba eZlevaT daukavSirdnen d.n.m-is matricas

anu moaxdinon hibridizacia (an `anilireba~ wyalbadis bmebis

meSveobiT) samizne sekvensis komplementarul adgils orive

bolodan, es procesi grZeldeba erTi an ramodenime wuTis

ganmavlobaSi.

3)Eelongacia-dagrZeleba, xdeba temperaturis momateba 720C-mde, 1

wuTis ganmavlobaSi, romelic saWiroa dnm polimerazas

funqcionirebisaTvis optimaluri temperaturis Sesaqmnelad. dnm

polimeraza sawyisi matricis komplementarulad praimerze axdens

dnm-is axali aslis agebas.

ciklirebisaTvis gamoviyeneT roSes firmis `laiT cikleri~

(LightCycler 2.0.) sistema, romelic Sedgeba xelawyosagan,

programisagan, reagentebisagan da dadgmis meTodikisagan.

myisieri PCR aRmoaCens zogadad brucelozis arsebobas an ar

arsebobas mocemuli nimuSis dnm-Si, Tumca SeuZlebelia

93

brucelas saxeobis dadgena, vinaidan myisieri PCR-is dros

gamoiyeneba genis is monakveTi, romelic brucelas yvela

saxeobisaTvis aris saerTo da mudmivi.

AMOS-PCR kvlevaSi gamoviyeneT 5 oligenukleotidis praimeris

koqteils, maT Soris Brucella erTi gvaris aRmomCeni praimeri IS711

sekvensisadmi da 4 praimeri specifiuri TiToeuli saxeobisaTvis.

aRniSnuli meTodi dafuZvnebulia genetikuri elementis IS711

saxeobis specifiur ganlagebaze qromosomaSi.

AMOS-PCR warmatebiT gamoiyeneba 498 kilofuZis mqone

nukleotidi B. abortus dnm-dan (msxvilfexa pirutyvis biovar) -1; 2

da 4-is diferencirebisaTvis.

IS711 praimeri: 5’_ TGCCGATCACTTAAGGGCCTTAT _3’

B. abortus praimeri: 5’ –GACGAACGGAATTTTTCCAATCCC _3’

cdebis procesSi gamokvlevas davuqvemdebareT soflis

meurneobis saministros laboratoriaSi, saqarTvelos sxvadasxva

raionebidan 2008-2010w.w. brucelozze gamosakvlevad Semosuli

mosaxleobis kerZo sakuTrebaSi arsebuli msxvilfexa pirutyvis

53 sisxlis Sratis, 1253 sisxlis, 577 rZis da 5 mogdebuli

nayofis nimuSebi. (cxr. 11). maT Soris eqstragirebuli 53 Sratis

nimuSidan miRebuli d.n.m-is nimuSi gamokvleul iqna myisier PCR-

ze, Cvens cdebSi B. abortus ocive Stamis mimarTebaSi Sedegi

aRmoCnda dadebiTi. saintereso Sedegi miviReT rogorc rZisa da

sisxlis ise mogdebuli nayofis nimuSebidan miRebuli

kulturebis eqstraqciis Sedegad, miRebuli d.n.m-i gamovikvlieT

94

myisieri PCR–iT, kulturas miviCnevdiT brucelad im SemTxvevaSi,

rodesac mrudis kveTa, romelic aisaxeba kompiuterSi

grafikulad iwyeboda me-11-13-e ciklidan me-40-e ciklis CaTvliT

(45-e cikli iTvleba sustad dadebiTad an saeWvod)

mocemuli suraTidan Cans, (suraTi 12) rom kveTis wertili (CP)

imyofeba me-12-13-e ciklebs Soris da mrudi Sesabamisad aris

sigmoiduri. aRniSnuli parametrebi miuTiTebs dadebiT Sedegze.

radgan myisieri PCR saSualebas iZleva aRmoaCinos brucelozis

arseboba zogadad. (dadebiTi an uaryofiTi). myisieri PCR-iT

miRebuli dadebiTi Sedegebi dadasturebul iqna AMOS PCR-iT.

(suraTi 13).

95

sur. 12. myisieri polimerazuli jaWvuri reaqciis dros dnm-is

amplifikaciis diagrama

96

sur. 13. AMOS PCR gelSi eleqtroforezi

1. B. abortus sakvlevi nimuSi.

2. B. abortus dadebiTi kontroli.

3. B. melitensis dadebiTi kontroli.

4. B. ovis dadebiTi kontroli.

1. 2. 3. 4.

d.n.m

lederi d.n.m

lederi

97

cxrili 11

polimerazuli jaWvuri reaqciiT gamokvleuli nimuSebi

# gamosakvlevi

masala nimuSebis raodenoba

Sedegebi

dadebiTi uaryofiTi

1 mogdebuli nayofi 5 1 4

2 rZe 577 16 565

3 sisxli 1253 3 1250

4 sisxlis Srati 53 31 22

98

4. miRebuli Sedegebis analizi

brucelozi gansakuTrebiT saSiS infeqciaTa Soris msoflio

masStabiT isev rCeba aqtualur problemad. cxovelTa dacvis

msoflio organizaciis (OIE) monacemebiT, dedamiwis xuTive

kontinentis 193 saxelmwifodan mxolod 38-ia Tavisufali

brucelozisagan; xuTSi infeqcia dafiqsirebulia gamonaklisi

SemTxvevebis saxiT. ormocdaaT qveyanaSi daavadebis erTeuli

SemTxvevebia aRwerili. enzootiuri gavrcelebis formiT

brucelozi gvxvdeba 38 saxelmwifoSi, xolo eqvsSi poulobs

farTe gavrcelebas; gansakuTrebiT xmelTaSua zRvis auzSi, mcire

aziaSi, arabeTis yureSi, meqsikaSi, centralur da samxreT

amerikaSi aRmosavleT aziaSi, afrikaSi.

brucelozi sazogadoebisaTvis ekonomikur da socialur

problemas warmoadgens. cxovelTa brucelozi farTodaa

gavrcelebuli msoflios bevr qveyanaSi, maT Soris

saqarTveloSic. is did ekonomiur zarals ayenebs mecxoveleobas,

xSiria adamianTa bruceloziT daavadebis SemTxvevebic.

bruceloziani adamiani Zalian Znelad inkurneba, xSirad igi rCeba

invalidad Sromisunarianobis dakargvis gamo. adamianTa

daavadebis aRkveTis saimedo garants warmoadgens am infeqciis

sruli da Seuqcevadi likvidacia cxovelebs Soris.

cxovelTa brucelozis likvidacia Zalze Znelia, rac aixsneba

rigi obieqturi mizezebiT. ZiriTadi am mizezTagan Semdegia:

paTogenobis farTe speqtri, rac saxeobrivi mravalferovnebiT

aris gamowveuli; daavadebis maRali kontagiozuroba; aRmZvrelis

99

unari, xangrZlivad iarsebon garemoSi (0,5-dan 5 Tvemde); cxovelur

produqtebsa da (10-dan 320 dRemde) da cxovelis organizmSi.

brucelebis gadacemis meqanizmsa da cxovelTa dainficirebis,

agreTve, infeqciis gavrcelebis gzebis mravalnairoba; xangrZlivi

inkubaciuri periodi; mxolod brucelozisaTvis damaxasiaTebeli

raime gansxvavebuli klinikuri niSnebis uqonloba. literaturaSi

aRwerilia avadmyofi cxovelis organizmidan aRmZvrelis Svidi da

ufro meti xnis ganmavlobaSi gamoyofis SemTxvevebi.

brusisa da bangis mier brucelas aRmoCenidan kolosaluri

raodenobis kvlevebia Catarebuli msoflios sxvadasxva qveynebSi,

Seswavlilia sakiTxTa farTe CamonaTvali, romelic Seexeba

cxovelTa brucelozs. miuxedavad diagnostikis, mkurnalobisa da

prevenciis mimarTebaSi arsebuli warmatebisa brucelozi kvlav

rCeba globaluri masStabis problemad jandacvisa da

veterinariisaTvis.

Cvens mier Catarebuli gamokvlevebi iTvaliswinebda

saqarTvelos sxvadasxva regionebSi brucelozis gavrcelebis

dinamikas da kartirebas. sisxlis Sratebis gamokvlevas roz-

bengal sinjiT da konkurentuli imunofermentuli analiziT, maTi

sadiagnostiko Rirebulebis dadgenas, paT. masalidan B. abortus

kulturaTa gamoyofas da etiologiuri rolis dadgenas

morfologiuri, kulturaluri niSan-Tvisebebis SeswavliT,

polimerazuli jaWvuri reaqciiT dnm-s skrinings da dnm.

analiziT B. abortus kulturaTaAidentificirebas.

statistikuri masalis gaanalizebiT brucelozi upiratesad

100

aRmosavleT saqarTveloSia gavrcelebuli, maT Soris daavadebis

xSiri SemTxvevebi modis kaxeTSi, qvemo-qarTlSi da mcxeTa-

mTianeTSi.

2008-2011w.w. msxvilfexa pirutyvis sisxlidan Cvens mier

gamoyofili Sratebis Seswavlam gviCvena, rom: 2008 wlis

ganmavlobaSi pirutyvis brucelozze gamosakvlevad Semosuli 3082

sisxlis Sratis sinjebidan, roz-bengal testiT dadebiTi

aRmoCnda 348, anu 11.30%, 2009 wels brucelozze gamokvleuli

msxvilfexa pirutyvis 6740 sisxlis Sratis sinjidan - 724, (10.74%).

xolo 2010 wels msxvilfexa pirutyvis 3654 sisxlis Sratidan -351

SemTxvevaSi, (9.61%). 2011 wels Semosuli da gamokvleuli 1699

sisxlis Sratidan, roz-bengal testiT dadebiTebma _ 227, anu

9,42% Seadgina.

yvela zemoT CamoTvlili roz-bengal dadebiTi Sratebis


Sedegebi identuria. rac miuTiTebs aRniSnuli meTodebis

sizustesa da specifiurobaze.

paralelurad 2008-2011w.w. furebis rZis gamokvlevas vaxdendiT

rZis rgoluri reaqciiT. gamokvlevebs davuqvemdebareT

saqarTvelos sxvadasxva raionebidan soflis meurneobis

saministros laboratoriaSi Semosuli mosaxleobis kerZo

sakuTrebaSi arsebuli 97 furis rZe.

maT Soris 2008w. 6 nimuSi, 2009w. _ 12, 2010w. _37, 2011w. _42 nimuSi.

gamokvleuli 97 rZis sinjidan 17 SemTxvevaSi rgoluri reaqcia

aRmoCnda dadebiTi, maT Soris 2008w. -1, (16.67%), 2009w. – 4, (33.34%),

101

2010w. -7, (18.925), 2011w. -5, (11.9%). aRsaniSnavia, rom Cvidmetive

dadebiTi SemTxveva aRiricxa mkveTr dadebiT reaqciad (+++),

rasac adasturebs sinjaraSi naRebis fenaSi tipuri lurj ferSi

SeRebili rgolis warmoqmna.

2010-2011w.w.-is ganmavlobaSi sakvalifikacio Temaze muSaobis

erT-erT ZiriTad mimarTulebas warmoadgenda msxvilfexa

pirutyvis paT. masalidan B. abortus kulturaTa gamoyofa da

identificireba. cdebis procesSi gamokvlevas davuqvemdebareT

saqarTvelos sami regionidan (qvemo qarTli, kaxeTi da imereTi)

2008-2010w.w. Semosuli 1253 sisxlis 577 rZis da 5 mogdebuli

nayofi, agreTve 53 Sratis nimuSi. sisxlis, rZis da mogdebuli

nayofis yvela nimuSi, Brucella-s gamosayofad vaxdendiT CaTesvas

xelovnur sakveb areebze, kerZod brucelas seleqtiur niadagze

da sisxlian agarze, nacxebis SeRebvas vaxdendiT gramiT da

kozlovskis meTodiT. gramis wesiT SeRebil preparatSi

mikroskopirebisas vnaxulobdiT wyvil-wyvilad ganlagebul

gramuaryofiT kokobacilebs (wiTeli feris), xolo kozlovskis

meTodiT SeRebil nacxebSi mowiTalo cal-calke an diplokokebad

dalagebul kokobaqteriebs. brucelebis bioqimiuri Tvisebebis

dadgena moicavda: oqsidazas, ureazas, katalazas testebs,

samSaqriani rkinis metabolizms, gogirdwyalbadis warmoqmnas.

B. abortus oqsidaza da ureaza dadebiTia. oqsidazas testiT xdeba

ferment citoqrom oqsidazas aRmoCena, is gamoyeneba gram-

uaryofiTi baqteriebis bioqimiuri testirebisaTvis. Cvens mier

gamoyofili B. abortus yvela izolati oqsidaza dadebiTia, rasac

102

adasturebs aRniSnuli testis Catarebisas lurji feris

warmoqmna.

ureazas testi gamoiyeneba mikrorganizmis mier ferment ureazas

saSualebiT Sardovanas hidrolizis TvisebaSi. Catarebuli

gamokvlevebiT davadgineT, rom B. abortus yvela Stami, iwvevs

vardisfer-wiTeli feris warmoqmnas, rac dadebiTi ureazas

testis maCvenebelia. Tumca feris Secvla ufro nela mimdinareobs

vidre brucela ureaza dadebiT sxva saxeobebSi.

B. abortus warmoqmnian gogirdwyalbads rasac adasturebs test-

stripze Savi Seferilobis miReba. brucelebi TSI-uaryofiTia.

brucelas saxeobaTa identifikaciisaTvis Cvens mier agreTve

gamoyenebul iqna Tioninisa da fuZe fuqsinis saRebavebic. B. abortus

izrdebian sakveb areSi fuZe fuqsinis arsebobisas, maSin rodesac

Tioninian sakveb areSi B. abortus ar izrdebian.

Brucella-as koliniebis morfologiis dasadastureblad

gamoviyeneT akriflavinis da kristal violetis testebi. B. abortus

mikroaerofilia, zrdisaTvis saWiroebs 10-15% CO2-s. aRniSnulis

gaTvaliswinebiT B. abortus kulturebis gamozrdas vaxdendiT 10-

15%CO2 -ian garemoSi.

sakvebisa da soflis meurneobis organizaciis (FAO), eqspertebis

rekomendaciebis gaTvaliswinebiT (1986), sxva kompleqsur

meTodebTan erTad B. abortus gamoyofili Stamebis

diferenciaciisaTvis gamoviyeneT baqteriofagis, mikrobiologiisa

da virusologiis intitutSi miRebuli Tb baqteriofagi.

103

aRniSnuli testi gamoiyeneba, rogorc damatebiTi Brucella-is

Stamebis identifikaciisaTvis.

Catarebulma gamokvlevebma gamoavlina B. abortus maRali

mgrZnobeloba Tb fagis mimarT. baqteriofagiT Seswavlili

kulturebis lizisis xarisxma 15 SemTxvevaSi Seadgina 4+, xolo 5

SemTxvevaSi 3+. miRebuli Sedegebi TvalnaTliv adasturebs, rom

gamoyofili Stamebi miekuTvneba tipur B. abortus saxeobas.

yvela zemoT CamoTvlili testirebis Sedegad miRebul iqna B.

abortus 20 saeWvo kultura, romelTa dadastureba moxda myisieri

PCR-iT da AMOS-PCR-iT. 31 Sratis nimuSi dadebiTi iyo

brucelozze.

brucelozis polimerazuli jaWvuri reaqciiT diagnostirebaze

mravali mkvlevari muSaobda mTels msoflioSi. (Fekete A. et. al., 1990;

Baily G.G. et. al., 1992; Bricker B.J. et. al.,1994; Leal-Klevezas D.S. et. al., 1995;

Romero C. et. al., 1995; Куличенко А.Н. 1995; Rijpens N.P. et.al. 1996;

Гаранина С.Б. 1996; Шумилов К.В.с соавт. 1996, 1998.).

bolo periodSi aRiniSneba dnm teqnologiis intensiuri

ganviTareba, romlis obieqturi mizezebi gaxda revoluciuri

aRmoCenebi molekulur biologiaSi da genur inJineriaSi. ramac

uzrunvelyo laboratoriuli meTodologiis axali mimarTulebis

Camoyalibeba –infeqciuri daavadebebis genodiagnostika.

genetikuri diagnostikis principi moicavs sxvadasxva meTodebiT

ganisazRvros specifiuri sakvlevi mikroorganizmis

nukleinismJavas ubani (dnm an rnm). genodiagnostikis ZiriTadi

104

mimarTulebebia dnm zondireba da nukleinis mJavebis

amplifikaciis meTodebi.

polimerazuli jaWvuri reaqciis dadgmis teqnika SedarebiT

xelmisawvdomia da amasTan erTad uzrunvelyofs sakmaod

maRalefeqturobas, specifiurobasa da analizis mgrZnobelobas.

gamokvlevis Sedegebis analizma gviCvena, rom sanitarul-

epidemiologiuri keTilsaimedoobis uzrunvelsayofad,

brucelozis sawinaaRmdegod RonisZiebebis efeqturad gatareba

saWiroebs daavadebuli cxovelebis droul gamovlinebas da

daavadebis swraf diagnostikas. serologiuri diagnostikisaTvis

erT-erTi yvelaze perspeqtiuli meTodia konkurentuli

imunofermentuli analizi da Tanamedrove molekulur-genetikuri

kvlevis meTodi – polimerazuli jaWvuri reaqcia.

105

5. d a s k v n e b i

1. brucelozi upiratesad aRmosavleT saqarTveloSia

gavrcelebuli 2,09%, dasavleT saqarTveloSi _ 1,5%-ia.

2. 2005-2011w.w. gamokvleuli 375426 suli msxvilfexa pirutyvidan

bruceloziT daavadebulTAricxvma roz-bengal sinjiT da

konkurentuli imunofermentuli analiziT 9493 (2,53%), xolo

2191 rZis nimuSis rgoluri reaqciiT gamokvleviT 103 (4,71%)

Seadgina.

3. saqarTveloSi msxvilfexa pirutyvis bruceloziT daavadebis

SedarebiT maRali intensivoba 2007, 2008, 2009 da 2010 wlebSi

aRiniSna. daavadebaze gansakuTrebiT arakeTilsaimedo

regionebia: kaxeTi, qvemo qarTli, mcxeTa-mTianeTi, imereTi.

4. roz-bengal sinji bruceloziT daavadebuli msxvilfexa

pirutyvis gamovlenis swrafi da specifikuri serologiuri

reaqciaa. misi saSualebiT brucelozze diagnozis dasma

umokles droSi 4 wuTSi xorcieldeba. roz-bengal sinjiT

gamovlenili Sedegebi saWiroebs imunofermentuli analiziT

Semdgom dazustebas.

5. roz-bengal sinjiT da konkurentuli imunofermentuli

analiziT gamovlenili brucelozze dadebiTi Sedegebi

Tanxvedrilia.

6. konkurentuli imunofermentuli analizi brucelozis

diagnostikis maRalmgrZnobiare da zedmiwevniT specifikuri

meTodia, romelic daavadebuli da vaqcinirebuli cxovelebis

diferencirebis saSualebas iZleva.

7. B. abortus gramuaryofiTi kokobacilaa, gramis da kozlovskis

106

wesiT SeRebvisas iRebebian Ria vardisferSi, agarze

warmoqmnian tipur S tipis koloniebs. seleqtiur farelis

sakveb agarze kultiviebisas mniSvnelovnad Cqardeba maTi

zdra-ganviTareba da pirveli generaciis miReba 48-72 saaTSia

SesaZlebeli.

8. paT. masalidan gamoyofili B. abortus kulturebi tipuri

brucelozis aRmZvelebia. isini xasiaTdebian katalazuri

aqtivobiT- oqsidaza da ureaza dadebiTia, gamoimuSaveben

gogirdwyalbads. TSI-uaryofiTebia. kultivirebisaTvis

saWiroebens 10-12% CO2-s.

9. polimerazuli jaWvuri reaqcia, msxvilfexa pirutyvSi

brucelozis mwvave da qronikuli formis diagnostikis

maRalmdrZnobiare, efeqturi da swrafi meTodia.

10. polimerazuli jaWvuri reaqciisaTvis damaxasiaTebelia

absoluturi specifikuroba, aprobirebuli meTodikiT

Catarebis SemTxvevaSi cru dadebiTi Sedegis miReba

gamoricxulia. AMOS PCR saSualebiT zustad xorcieldeba

brucelebis saxeobaTa diferencireba, rac daavadebis

sawinaaRmdego RonisZiebaTa droulad da efeqturad

gatarebis safuZvelia.

107

6. praqtikuli winadadebebi

1. brucelozis serodiagnostikisaTvis roz-bengal testis

dasadastureblad imunofermentul analizs, romelic

mniSvnelovnad swrafi, efeqturi da maRalmgrZnobiarea.

2. B. aborus Stamebis kultivirebisa da identifikaciisaTvis:

seleqtiuri farelis agaris, ureazas, oqsidazas, katalazuri

aqtivobis dadgenis, gogirwyalbadis warmoqmnis testebsa da Tb

fagiT tipirebis gamoyenebas.

3. raionul da savele laboratoriebSi molekulur-biologiuri

kvlevis swraf da efeqtur meTods-polimerazul jaWvur reaqcias

(p.j.r), romelic saSualebas iZleva Catardes brucelaTa

genetikuri analizi.

4. brucelaTa saxeobebis, biotipebisa da Stamebis swrafi

diferencirebisaTvis AMOS PCR testis farTed gamoyenebas.

108

7. gamoyenebuli literatura

1. babakiSvili j., mamaiaSvili m., cxakaia T., kereseliZe m.

,,cxovelTa infeqciuri daavadebebi”. Tbilisi 2005, gv.53-65.

2. gamyreliZe i., berianiZe e., babakiSvili T. `rZis gamokvleva

brucelozze.~ agraruli mecnierebis problemebi, samecniero

SromaTa krebuli, XXXI Tbilisi. 2005w. gv.184-186.

3. koliakovi i. `veterinaruli mikrobiologia~ Tbilisi 1987w.

gv.73-82.

4. lomineiSvili m., gavaSeli T. ,,brucelozi da misi sawinaaRmdego

RonisZiebebi” Tbilisi. 1995.

5. naTiZe m., WaniSvili T., gomareli g. `baqteriofagi~. Tbilisi

1989w.

6. naTiZe m. `sasoflo-sameurneo cxovelebis infeqciur

daavadebaTa serologiuri diagnostika.~ Tbilsi 1989w.

7. naTiZe m., gabisonia t., rigvava s. `zogadi virusologia~.

Tbilisi 1996w. gv. 147-148.

8. naTiZe m., gabisonia t., naTiZe T., rigvava s. `baqteriologiuri

gamokvlevebis safuZvlebi saveterinaro medicinaSi~. Tbilisi

2005w. gv. 98-102.

9. naTiZe m., giorgaZe m. ,,brucelozis aRmZvreli”. meToduri

miTiTeba saveterinaro medicinisa da mecxoveleobis produqtebis

warmoebisa da gadamuSavebis fakultetis studentebisaTvis.

Tbilisi 2006w.

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