Quy Trinh Ky Thuat Chuyen Nganh Hoa Sinh
description
Transcript of Quy Trinh Ky Thuat Chuyen Nganh Hoa Sinh
-
1
B Y T
S: 320 /Q-BYT
CNG HA X HI CH NGHA VIT NAM
c lp - T do - Hnh phc
H Ni, ngy 23 thng 01 nm 2014
QUYT NH V vic ban hnh ti liu
Hng dn quy trnh k thut chuyn ngnh Ha sinh
B TRNG B Y T
Cn c Lut khm bnh, cha bnh nm 2009;
Cn c Ngh nh s 63/2012/N-CP ngy 31/8/2012 ca Chnh Ph quy nh chc nng, nhim v, quyn hn v c cu t chc ca B Y t;
Xt Bin bn hp ca Hi ng nghim thu Hng dn Quy trnh k thut khm bnh, cha bnh chuyn ngnh Ha sinh ca B Y t;
Theo ngh ca Cc trng Cc Qun l Khm, cha bnh,
QUYT NH:
iu 1. Ban hnh km theo Quyt nh ny ti liu Hng dn quy trnh k thut chuyn ngnh Ha sinh, gm 220 quy trnh k thut.
iu 2. Ti liu Hng dn quy trnh k thut chuyn ngnh Ha sinh ban hnh km theo Quyt nh ny c p dng ti cc c s khm bnh, cha bnh.
Cn c vo ti liu hng dn ny v iu kin c th ca n v, Gim c c s khm bnh, cha bnh xy dng v ban hnh ti liu Hng dn Quy trnh k thut Ha sinh ph hp thc hin ti n v.
iu 3. Quyt nh ny c hiu lc k t ngy k ban hnh.
iu 4. Cc ng, b: Chnh Vn phng B, Chnh Thanh tra B, Cc trng Cc Qun l Khm, cha bnh, Cc trng v V trng cc Cc, V thuc B Y t, Gim c cc bnh vin, vin c ging bnh trc thuc B Y t, Gim c S Y t cc tnh, thnh ph trc thuc trung ng, Th trng Y t cc B, Ngnh v Th trng cc n v c lin quan chu trch nhim thi hnh Quyt nh ny./.
Ni nhn: - Nh iu 4; - B trng B Y t ( b/c); - Cc Th trng BYT; - Bo him X hi Vit Nam ( phi hp); - Cng thng tin in t BYT; - Website Cc KCB; - Lu VT, KCB.
KT. B TRNG TH TRNG
k
Nguyn Th Xuyn
-
2
B Y T CNG HA X HI CH NGHA VIT NAM
c lp - T do- Hnh Phc
DANH MC QUY TRNH K THUT CHUYN NGNH HA SINH
(Ban hnh km theo Quyt nh s: 320/Q-BYT ngy23 thng 01 nm 2014 ca B trng B Y t)
TT TN QUY TRNH K THUT
A. MU
1 o hot ACP (Phosphatase Acid)
2 nh lng ACTH
3 nh lng Acid Uric
4 nh lng ADH (Anti Diuretic Hormone)
5 nh lng Adiponectin
6 nh lng Aldosteron
7 nh lng Albumin
8 nh lng Alpha1 Antitrypsin
9 o hot ALP (Alkalin Phosphatase)
10 o hot Amylase
11 nh lng Amoniac ( NH3)
12 nh lng AMH ( Anti- Mullerian Hormone)
13 nh lng Anti CCP
14 nh lng Anti-Tg (Antibody- Thyroglobulin)
15 nh lng Anti - TPO (Anti- thyroid Peroxidase antibodies)
16 nh lng Apo A1 (Apolipoprotein A1)
17 nh lng Apo B (Apolipoprotein B)
18 nh lng AFP (Alpha Fetoproteine)
19 o hot ALT (GPT)
20 o hot AST (GOT)
21 nh lng 1 Acid Glycoprotein
22 nh lng 2 microglobulin
23 nh lng Beta Crosslap
24 nh lng hCG (Beta human Chorionic gonadotropins)
25 nh lng Bilirubin trc tip
26 nh lng Bilirubin gin tip
27 nh lng Bilirubin ton phn
28 nh lng BNP (B- Type Natriuretic Peptide)
29 nh lng Calci ton phn
30 nh lng Calci ion ho
31 nh lng canci ion ha bng in cc chn lc
32 nh lng CA 125 (cancer antigen 125)
-
3
33 nh lng CA 19 - 9 (carbohydrate antigen 19-9)
34 nh lng CA 15 - 3 (cancer antigen 15- 3)
35 nh lng CA 72 - 4 (cancer antigen 72- 4)
36 nh lng Calcitonin
37 nh lng Carbamazepin
38 nh lng Ceruloplasmin
39 nh lng CEA (carcino embryonic antigen)
40 o hot Cholinesterase (ChE)
41 nh lng Cholesterol ton phn
42 o hot CK (Creatine kinase)
43 o hot CK-MB (Isozym MB of Creatine kinase)
44 nh lng CK-MB mass
45 nh lng C-Peptid
46 nh lng Cortisol
47 nh lng Cystatine C
48 nh lng b th C3
49 nh lng b th C4
50 nh lng CRP hs (C-reactive protein high sesitivity)
51 nh lng Creatinin
52 nh lng Cyfra 21- 1
53 nh lng cyclosphorin
54 nh lng D-Dimer
55 nh lng 25OH Vitamin D (D3)
56 nh lng Digoxin
57 nh lng Digitoxin
58 nh lng cc cht in gii (Na, K, Cl)
59 nh lng FABD (Fatty acid binding protein)
60 nh lng Ethanol (cn)
61 nh lng Estradiol
62 nh lng E3 khng lin hp (Unconjugated Estriol)
63 nh lng Ferritin
64 nh lng Fructosamin
65 nh lng FSH (Follicular stimulating hormone)
66 nh lng free HCG (Free Beta Human chorionic gonadotropin)
67 nh lng Folate
68 nh lng FT3 (Free Triiodothyronine)
69 nh lng FT4 (Free thyroxine)
70 nh lng Galectin 3
71 nh lng Gastrin
72 o hot G6PD (Glucose -6 phosphat dehydrogenase)
-
4
73 nh lng GH (Growth Hormone)
74 o hot GLDH (Glutamat dehydrogenase)
75 nh lng Glucose
76 nh lng Globulin
77 o hot GGT (Gama Glutamyl Transferase)
78 nh lng GLP-1
79 nh lng Gentamicin
80 nh lng Haptoglobulin
81 nh lng HBs g (HBs g Quantitative) ( CMI / ECLI )
82 o hot HBDH (Hydroxy butyrat dehydrogenase)
83 nh lng HbA1c
84 nh lng HDL-C (High density lipoprotein Cholesterol)
85 nh lng HE4
86 nh lng Homocystein
87 nh lng IL-1 (Interleukin 1)
88 nh lng IL -1 (Interleukin 1)
89 nh lng IL-6 ( Interleukin 6)
90 nh lng IL-8 (Interleukin 8)
91 nh lng IL-10 (Interleukin 10)
92 nh lng IgE (bng phng php ELIS )
93 nh lng IgE (Immunoglobuline E)
94 nh lng IgA (Immunoglobuline A)
95 nh lng IgG (Immunoglobuline G)
96 nh lng IgM (Immunoglobuline M)
97 nh lng IGFBP-3 ( Insulin like growth factor binding protein 3)
98 nh lng Insulin
99 in di Isozym LDH
100 nh lng IMA (Ischemia Modified Albumin)
101 nh lng Kappa
102 nh lng Kappa t do (Free kappa)
103 Xt nghim Kh mu
104 nh lng Lactat (Acid Lactic)
105 nh lng Lambda
106 nh lng Lambda t do (Free Lambda)
107 nh lng Leptin human
108 in di LDL/HDL cholesterol
109 o hot Lipase
110 nh lng LH (Luteinizing hormone)
111 o hot LDH ( Lactat dehydrogenase)
112 nh lng LDL - C (Low density lipoprotein Cholesterol)
-
5
113 in di Lipoprotein
114 nh lng Lp-PLA2 (Lipoprotein Associated Phospholipase A2)
115 nh lng Malondialdehyd (MDA)
116 o hot MPO
117 nh lng Myoglobin
118 nh lng Mg
119 nh lng N-MID Osteocalcin
120 nh lng NSE (Neuron Specific Enolase)
121 nh lng NT-proBNP
122 o hot P-Amylase
123 nh lng PAPP-A
124 nh lng Pepsinogen I
125 nh lng Pepsinogen II
126 nh lng Phenobarbital
127 nh lng Phenytoin
128 nh lng Phospho
129 nh lng Pre-albumin
130 nh tnh Pro-calcitonin
131 nh lng Prolactin
132 in di protein
133 nh lng Protein ton phn
134 nh lng Progesteron
135 nh lng Procainnamid
136 nh lng protein S100
137 nh lng Pro-GRP ( Pro- Gastrin-releasing peptide)
138 nh lng PSA t do (Free prostate-specific antigen)
139 nh lng PSA ton phn (Total prostate-specific antigen)
140 nh lng PTH (Parathyroid hormon)
141 nh lng Renin activity
142 nh lng RF (Reumatoid factor)
143 nh lng St
144 nh lng SCC (squamous cell carcinoma antigen)
145 nh lng SHBG (Sex hormon binding globulin)
146 nh lng Sperm Antibody
147 nh lng T3 (Tri iodothyronine)
148 nh lng T4 (Thyroxine)
149 nh lng s TfR (solube transferin receptor)
150 nh lng Tacrolimus
151 nh lng Testosterol
152 nh lng TGF 1( Transforming Growth Factor Beta 1)
-
6
153 nh lng TGF 2( Transforming Growth Factor Beta 2)
154 nh lng Tg (Thyroglobulin)
155 nh lng Theophylline
156 nh lng TRAb (TSH Receptor Antibodies)
157 nh lng Transferin
158 nh lng Triglycerid
159 nh lng Troponin T
160 nh lng Troponin T hs
161 nh lng Troponin I
162 nh lng TSH (Thyroid Stimulating hormone)
163 nh lng Tobramycin
164 nh lng Total p1NP
165 nh lng T-uptake
166 nh lng Ur
167 nh lng Valproic acid
168 nh lng Vancomycin
169 nh lng Vitamin B12
170 nh lng PLGF (Placental Growth Factor- yu t tn to mch mu)
171 nh lng sFlt-1 (yu t khng tn to mch mu)
B. NC TIU
172 nh lng cc cht in gii
173 nh tnh Amphetamin
174 nh lng Amphetamin
175 o hot Amylase
176 nh lng axit uric
177 nh lng Barbiturates
178 nh lng Benzodiazepin
179 nh tnh HCG
180 nh lng Canxi
181 nh lng Catecholamin
182 nh lng Cocain
183 nh lng Cortisol
184 nh lng Creatinin
185 nh lng dng chp
186 nh tnh dng chp
187 nh lng Glucose
188 nh tnh Marijuana
189 nh lng MAU
190 nh lng Mathadon
-
7
191 nh lng NGAL
192 nh lng opiat
193 nh tnh Morphin
194 nh lng Phospho
195 nh tnh phospho hu c
196 nh tnh Porphyrin
197 in di protein
198 nh lng Protein
199 nh tnh Protein Bence jones
200 nh tnh Rotundin
201 nh lng THC (Canabionids)
202 nh lng Ure
203 Tng phn tch nc tiu (bng my t ng)
C. DCH NO TU
204 nh lng Clo
205 nh lng Glucose
206 Phn ng Pandy
207 nh lng Protein
D. THY DCH MT
208 nh lng Albumin
209 nh lng Globulin
E. DCH CHC D (Dch mng bng, mng phi, mng tim)
210 o hot Amylase
211 nh lng Bilirubin ton phn
212 nh lng Cholesterol ton phn
213 nh lng Creatinin
214 nh lng Glucose
215 o hot LDH
216 nh lng Protein ton phn
217 Phn ng Rivalta
218 nh lng Triglycerid
219 o t trng dch chc d
220 nh lng Ure
(Tng s 220 quy trnh k thut) KT. B TRNG
TH TRNG
Nguyn Th Xuyn
-
8
MC LC
A. MU
1. o hot CP (Phosphatase cid) 9
2. nh lng CTH 12
3. nh lng Acid Uric 17
4. nh lng DH ( nti Diuretic Hormone) 20
5. nh lng diponectin 24
6. nh lng ldosteron 29
7. nh lng Albumin 36
8. nh lng lpha1 ntitrypsin 39
9. o hot LP ( lkalin Phosphatase) 42
10. o hot mylase 45
11. nh lng moniac ( NH3) 48
12. nh lng MH ( nti- Mullerian Hormone) 51
13. nh lng nti CCP 57
14. nh lng nti-Tg (Antibody- Thyroglobulin) 60
15. nh lng nti - TPO (Anti- thyroid Peroxidase antibodies) 63
16. nh lng Apo A1 (Apolipoprotein A1) 66
17. nh lng po B ( polipoprotein B) 69
18. nh lng FP ( lpha Fetoproteine) 72
19. o hot LT (GPT) 75
20. o hot ST (GOT) 79
21. nh lng 1 Acid Glycoprotein 83
22. nh lng 2 microglobulin 86
23. nh lng Beta Crosslap 89
24. nh lng hCG (Beta human Chorionic gonadotropins) 94
25. nh lng Bilirubin trc tip 97
26. nh lng Bilirubin gin tip 100
-
9
27. nh lng Bilirubin ton phn 102
28. nh lng BNP (B- Type Natriuretic Peptide) 105
29. nh lng Calci ton phn 109
30. nh lng Calci ion ho 112
31. nh lng canci ion ha bng in cc chn lc 114
32. nh lng CA 125 (cancer antigen 125) 116
33. nh lng CA 19 - 9 (carbohydrate antigen 19-9) 119
34. nh lng CA 15 - 3 (cancer antigen 15- 3) 122
35. nh lng CA 72 - 4 (cancer antigen 72- 4) 125
36. nh lng Calcitonin 128
37. nh lng Carbamazepin 132
38. nh lng Ceruloplasmin 135
39. nh lng CEA (carcino embryonic antigen) 138
40. o hot Cholinesterase (ChE) 141
41. nh lng Cholesterol ton phn 145
42. o hot CK (Creatine kinase) 148
43. o hot CK-MB (Isozym MB of Creatine kinase) 152
44. nh lng CK-MB mass 155
45. nh lng C-Peptid 159
46. nh lng Cortisol 163
47. nh lng Cystatine C 168
48. nh lng b th C3 172
49. nh lng b th C4 175
50. nh lng CRP hs (C-reactive protein high sesitivity) 178
51. nh lng Creatinin 181
52. nh lng Cyfra 21- 1 184
53. nh lng cyclosphorin 187
54. nh lng D-Dimer 192
55. nh lng 25OH Vitamin D (D3) 196
56. nh lng Digoxin 200
-
10
57. nh lng Digitoxin 203
58. nh lng cc cht in gii (Na, K, Cl) 205
59. nh lng F BD (Fatty acid binding protein) 208
60. nh lng Ethanol (cn) 213
61. nh lng Estradiol 216
62. nh lng E3 khng lin hp (Unconjugated Estriol) 219
63. nh lng Ferritin 224
64. nh lng Fructosamin 227
65. nh lng FSH (Follicular stimulating hormone) 230
66. nh lng free HCG (Free Beta Human chorionic gonadotropin)
233
67. nh lng Folate 237
68. nh lng FT3 (Free Triiodothyronine) 241
69. nh lng FT4 (Free thyroxine) 244
70. nh lng Galectin 3 247
71. nh lng Gastrin 250
72. o hot G6PD (Glucose -6 phosphat dehydrogenase) 255
73. nh lng GH (Growth Hormone) 258
74. o hot GLDH (Glutamat dehydrogenase) 264
75. nh lng Glucose 266
76. nh lng Globulin 269
77. o hot GGT (Gama Glutamyl Transferase) 271
78. nh lng GLP-1 275
79. nh lng Gentamicin 280
80. nh lng Haptoglobulin 283
81. nh lng HBs g (HBs g Quantitative) ( CMI / ECLI ) 286
82. o hot HBDH (Hydroxy butyrat dehydrogenase) 290
83. nh lng Hb 1c 292
84. nh lng HDL-C (High density lipoprotein Cholesterol) 296
85. nh lng HE4 299
-
11
86. nh lng Homocystein 303
87. nh lng IL-1 (Interleukin 1) 306
88. nh lng IL -1 (Interleukin 1) 309
89. nh lng IL-6 ( Interleukin 6) 312
90. nh lng IL-8 (Interleukin 8) 316
91. nh lng IL-10 (Interleukin 10) 319
92. nh lng IgE (bng phng php ELIS ) 322
93. nh lng IgE (Immunoglobuline E) 326
94. nh lng Ig (Immunoglobuline ) 329
95. nh lng IgG (Immunoglobuline G) 332
96. nh lng IgM (Immunoglobuline M) 335
97. nh lng IGFBP-3 ( Insulin like growth factor binding protein 3)
338
98. nh lng Insulin 344
99. in di Isozym LDH 348
100. nh lng IM (Ischemia Modified lbumin) 353
101. nh lng Kappa 358
102. nh lng Kappa t do (Free kappa) 361
103. Xt nghim Kh mu 365
104. nh lng Lactat ( cid Lactic) 369
105. nh lng Lambda 372
106. nh lng Lambda t do (Free Lambda) 375
107. nh lng Leptin human 379
108. in di LDL/HDL cholesterol 383
109. o hot Lipase 388
110. nh lng LH (Luteinizing hormone) 391
111. o hot LDH ( Lactat dehydrogenase) 394
112. nh lng LDL - C (Low density lipoprotein Cholesterol) 397
113. in di Lipoprotein 400
114. nh lng Lp-PLA2 (Lipoprotein Associated Phospholipase 403
-
12
A2)
115. nh lng Malondialdehyd (MD ) 408
116. o hot MPO 412
117. nh lng Myoglobin 415
118. nh lng Mg 418
119. nh lng N-MID Osteocalcin 422
120. nh lng NSE (Neuron Specific Enolase) 426
121. nh lng NT-proBNP 429
122. o hot P-Amylase 434
123. nh lng P PP-A 437
124. nh lng Pepsinogen I 442
125. nh lng Pepsinogen II 447
126. nh lng Phenobarbital 452
127. nh lng Phenytoin 455
128. nh lng Phospho 458
129. nh lng Pre-albumin 461
130. nh tnh Pro-calcitonin 465
131. nh lng Prolactin 468
132. in di protein 471
133. nh lng Protein ton phn 474
134. nh lng Progesteron 478
135. nh lng Procainnamid 481
136. nh lng protein S100 484
137. nh lng Pro-GRP ( Pro- Gastrin-releasing peptide) 487
138. nh lng PS t do (Free prostate-specific antigen) 492
139. nh lng PS ton phn (Total prostate-specific antigen) 495
140. nh lng PTH (Parathyroid hormon) 498
141. nh lng Renin activity 501
142. nh lng RF (Reumatoid factor) 506
143. nh lng St 509
-
13
144. nh lng SCC (squamous cell carcinoma antigen) 512
145. nh lng SHBG (Sex hormon binding globulin) 517
146. nh lng Sperm ntibody 521
147. nh lng T3 (Tri iodothyronine) 525
148. nh lng T4 (Thyroxine) 528
149. nh lng s TfR (solube transferin receptor) 531
150. nh lng Tacrolimus 534
151. nh lng Testosterol 538
152. nh lng TGF 1( Transforming Growth Factor Beta 1) 541
153. nh lng TGF 2( Transforming Growth Factor Beta 2) 546
154. nh lng Tg (Thyroglobulin) 551
155. nh lng Theophylline 554
156. nh lng TR b (TSH Receptor ntibodies) 557
157. nh lng Transferin 562
158. nh lng Triglycerid 565
159. nh lng Troponin T 569
160. nh lng Troponin T hs 572
161. nh lng Troponin I 576
162. nh lng TSH (Thyroid Stimulating hormone) 579
163. nh lng Tobramycin 582
164. nh lng Total p1NP 585
165. nh lng T-uptake 589
166. nh lng Ur 592
167. nh lng Valproic acid 595
168. nh lng Vancomycin 598
169. nh lng Vitamin B12 601
170. nh lng PLGF (Placental Growth Factor- yu t tn to mch mu)
605
171. nh lng sFlt-1 (yu t khng tn to mch mu) 610
B NC TIU
-
14
172. nh lng cc cht in gii 614
173. nh tnh mphetamin 618
174. nh lng mphetamin 621
175. o hot mylase 624
176. nh lng axit uric 626
177. nh lng Barbiturates 630
178. nh lng Benzodiazepin 633
179. nh tnh HCG 636
180. nh lng Canxi 638
181. nh lng Catecholamin 641
182. nh lng Cocain 644
183. nh lng Cortisol 647
184. nh lng Creatinin 650
185. nh lng dng chp 653
186. nh tnh dng chp 656
187. nh lng Glucose 659
188. nh tnh Marijuana 661
189. nh lng M U 664
190. nh lng Mathadon 667
191. nh lng NG L 670
192. nh lng opiat 674
193. nh tnh Morphin 676
194. nh lng Phospho 679
195. nh tnh phospho hu c 682
196. nh tnh Porphyrin 687
197. in di protein 690
198. nh lng Protein 693
199. nh tnh Protein Bence jones 696
200. nh tnh Rotundin 698
201. nh lng THC (Canabionids) 703
-
15
202. nh lng Ure 706
203. Tng phn tch nc tiu (bng my t ng) 709
C. DCH NO TU
204. nh lng Clo 713
205. nh lng Glucose 716
206. Phn ng Pandy 719
207. nh lng Protein 721
D. THY DCH MT
208. nh lng lbumin 724
209. nh lng Globulin 726
E. DCH CHC D (Dch mng bng, mng phi, mng tim)
210. o hot mylase 728
211. nh lng Bilirubin ton phn 731
212. nh lng Cholesterol ton phn 733
213. nh lng Creatinin 736
214. nh lng Glucose 740
215. o hot LDH 743
216. nh lng Protein ton phn 745
217. Phn ng Rivalta 748
218. nh lng Triglycerid 751
219. o t trng dch chc d 754
220. nh lng Ure 756
-
16
A. MU
1. O HOT ACP
(Phosphatase acid)
I. NGUYN L
CP xc tc phn ng thu phn 1-naphthyl phosphat pH 4.5- 6 gii phng phosphate v c v 1-naphtanol. 1-naphtanol phn ng vi FRTR to sn phm mu azo. Hot enzym c tnh ton bng s thay i mt quang ti bc sng
II. CHUN B
1. Ngi thc hin K thut vin ca phng xt nghim c nhim v nhn v kim tra cht lng ca
mu bnh phm bng cch i chiu vi cc tiu chun loi b v thc hin phn tch theo phng php c xc nh.
2. Phng tin, ha cht
- My ha sinh t ng U2700
- My ly tm
- Ha cht lm xt nghim CP (hng Olympus )
- Huyt thanh kim tra mc 1 (QC mc bnh thng)
- Huyt thanh kim tra mc 2 (QC mc bnh l)
- Chun
- Cht n nh CP
- Nc ct
3. Ngi bnh
Khng thm khm tuyn tin lit trc khi ly mu mu v c th lm tng hot ACP.
4. Phiu xt nghim
Ghi y thng tin cn thit: h tn ngi bnh, tui, m s ngi bnh, khoa phng, tn xt nghim ch nh, khong tham chiu, bc s ch nh xt nghim, ngy gi ly mu, ngi ly mu, ngy gi nhn mu bnh phm, ngi nhn mu.
III. CC BC TIN HNH
1. Ly bnh phm
-
17
Huyt thanh, khng dng huyt tng.
2.Tin hnh k thut
- Ly tm ng mu trong 5 pht vi vn tc 3000 vng/ pht.
- Tch huyt thanh v thm mt git cht n nh ACP - t ng bnh phm vo v tr trn khay cha mu. - Vn hnh my theo hng dn trong ti liu hng dn s dng my. - My s t ng in ra kt qu sau khi hon tt qu trnh phn tch. - Kim sot cht lng:
Hng ngy: Chy 2 mc kim QC tra cht lng hng ngy vo bui sng v t nht sau mi 8 ting. Tt c cc kt qu kim tra cht lng phi c ghi li trong bng theo di QC. Ch thng bo kt qu xt nghim nu c hai mc QC nm trong khong cho php.
nh k : Chun li v chy 2 mc QC sau khi thay l thuc th mi hoc sau khi bo dng, sa cha my do s c, thay th trang thit b phn tch quan trng. Ghi li kt qu vo bng theo di chun my XN.
IV. NHN NH KT QU 1. Gi tr tham chiu
Hot CP: Ngi ln 4,8- 13,5 U/L 37oC 2. ngha lm sng CP tng cao l ch im cho carcinoma tin lit tuyn, bnh xng hoc bnh ca h vng ni m.
V. NHNG SAI ST V X TR.
- V CP khng bn pH trn 7, ngay sau khi ly mu phi tch huyt thanh, cho thm 20 l/ ml huyt thanh dung dch acid acetic 10% hoc sodium bisulfat.
- Mu khng acid ho hot gim 20% trong vng 3gi. Mu acid ho n nh 24 gi nhit phng v n nh 7 ngy khi bo qun ti 2 n 8oC
-
18
2. NH LNG ACTH MU
(Adrenocorticotropic hormone)
I. NGUYN L
ACTH hoc corticotropin l mt hormone peptide gm 39 amino acid, c sn xut bi thy trc tuyn yn. ACTH kch thch s hnh thnh v s bi tit glucocorticoid (c bit l cortisol) ca vng v ca tuyn thng thn.
CTH c nh lng bng phng php min dch kiu sandwich s dng cng ngh in ha pht quang (ECLIA). Tng thi gian phn tch mt mu l 18 pht.
+ Giai on u tin: To thnh phc hp sandwich gm 3 thnh phn:
Mu bnh phm (huyt thanh, huyt tng) kp gia khng th n dng c hiu khng CTH gn biotin v mt khng th n dng c hiu, khng ACTH gn vi ruthenium. Ba thnh phn to thnh mt phc hp min dch kiu bnh sandwich
+ Giai on th hai: Sau khi cho thm cc vi ht c bao ph bi Streptavidin. Phc hp c gn kt vo pha rn do s tng tc gia biotin v streptavidin.
+ Phc hp phn ng c a vo bung o. Ti y cc vi ht (microparticles) c gi li bng t tnh trn b mt in cc. Nhng cht tha c ra i bng procell. Dng mt dng in mt chiu tc ng vo in cc nhm kch thch pht quang v cng tn hiu nh sng pht ra c th o c bng b phn nhn quang.
+ Kt qu c tnh ton da vo ng cong chun thu c bng cch chun 2 im v ng cong gc c cung cp t nh sn xut. Nng cht cn nh lng t l thun vi cng nh sng thu c.
II. CHUN B
1. Ngi thc hin
01 Cn b i hc chuyn ngnh Ha sinh min dch v 01 K thut vin
2. Phng tin, ha cht
2.1. Phng tin
- Cc my c th phn tch: Elecsys 1010, Elecsys 2010, Modular analytics e 170, cobas e 411, cobas e 601 v mt s my khc
- My ly tm
- T lnh bo qun ha cht v bo qun QC, mu bnh phm
-
19
- Pipet cc loi
- ng nghim, u cn xanh v vng
- Gi ng ng nghim
2.2.Ha cht
- L 1 (M) np trong: Streptavidin-coated microparticles, th tch 6,5 mL: (c cha Streptavidin-coated microparticles 0,72 mg/mL) v cht bo qun.
- L 2 (R1) - np mu xm: nti-ACTH- b~biotin, th tch 8 mL (c cha Biotinylated monoclonal anti-ACTH antibody (mouse) 0.3 mg/L); dung dch m MES (2-morpholino-ethane sulfonic acid) 50 mmol/L, pH 6,2 v cht bo qun.
- L 3 (R2)- np mu en: nti-ACTH- b~Ru(bpy), th tch 8 mL (c cha Monoclonal anti- CTH antibody (mouse) gn vi phc hp ruthenium 0.3 mg/L)
- Dung dch m MES (2-morpholino-ethane sulfonic acid) 50 mmol/L, pH 6.2 v cht bo qun.
Thuc th c bo qun nhit 2- 80C, c th n nh n thi hn ghi trn hp. Thuc th sau khi m np bo qun c 12 tun 2-8oC. Nu trn my (khng tt my) c th c 4 tun.
+ Procell; Clean cell
+ Dung dch chun
+ Quality control (QC): gm 3 mc: level 1, 2 v 3
2.3. Cc dng c tiu hao khc
- ng nghim
- Gng tay
- Bng , cn st trng, bm tim hoc kim ly mu
3. Ngi bnh
Cn gii thch cho ngi bnh v ngi nh ngi bnh v mc ch ca xt nghim
Ngi bnh cn phi hp ly mu theo ng yu cu v thi gian v s lng
4. Phiu xt nghim
C phiu xt nghim ghi r yu cu xt nghim. Trn phiu cn ghi r thi gian ly mu trong ngy trn ng (mu 1, mu 2).
III. CC BC TIN HNH
1. Ly bnh phm
-
20
Mu mu c ly vo ng chng ng EDT . Mu c vo khay , chuyn xung PXN ngay. Khi tch huyt tng s dng ly tm lnh. Mu cn c phn tch ngay. C th n nh 2h 220C. Nu khng phi lu gi nhit -200C (ch c ng 1 ln)
m bo mu ngi bnh, dung dch chun v QC phi nhit t 22-250C trc khi phn tch
2. Tin hnh k thut
2.1. Chun b my phn tch
Dng ng chun
Phn tch QC: c 3 level. Khi QC t tin hnh phn tch mu
2.2. Phn tch mu
Mu bnh phm nn c tin hnh phn tch trong vng 2h
Mu sau khi ly tm c chuyn vo khay ng bnh phm
nh s (hoc ID ca ngi bnh); chn test v my s t ng phn tch.
IV. NHN NH KT QU
Tr s bnh thng: Bui sng < 18 pmol/L (< 80 pg/mL)
Bui chiu: < 11 pmol/L ( 428 mmol/L (>25 mg / dL); mu b huyt tn nhng Hb >0,25 mmol/L (>0,4 g/dL), Triglycerid mu >1500 mg / dL), v biotin > 246 nmol/L (> 60 ng/mL); yu t dng thp (RF> 400 IU/mL.
- Ngi bnh ang iu tr liu cao biotin (> 5 mg/ngy), nn ly mu mu cch xa thi gian ung thuc ln cui cng 8gi.
Cc yu t Hu qu X tr
Ngi bnh b sang chn (stress), h glucose mu, ph n ang mang thai
C th lm tng kt qu Ch khi nhn nh kt qu
Cc thuc ang s dng: aminoglutethimide,
amphetamines, estrogen,
C th lm tng kt qu C th ngng thuc trc khi c ch nh xt nghim
-
21
ethanol, insulin
Thuc th, dung dch chun v dung dch QC, bo qun nhit khng ng quy nh v b bt
nh hng n kt qu xt nghim
Bo qun thuc th, dung dch chun, dung dch QC ng nhit quy nh, Trnh gy bt
-
22
3. NH LNG ACID URIC
I. NGUYN L
Acid Uric l sn phm chuyn ha cui cng ca base c nit nhn purin
Acid Uric mu c nh lng theo phng php enzyme so mu
Uric Acid + 2H2O + O2 Uricase Allatoin + CO2 + H2O2
peroxidase
2 H2O2 + H+ + TOOS
a + 4-aminophenazone hp cht mu + 4H2O
Sn phm mu c o bc sng 546nm.
II. CHUN B
1. Ngi thc hin
Bc s hoc k thut vin c o to chuyn ngnh Ha sinh
2. Phng tin, ha cht
- My mc: h thng my sinh ha
- Thuc th: sn sng s dng.
R 1: buffer, TOOS ...
R 2: Uricase, POD, 4-AAP...
Bo qun 2-80C n khi ht hn s dng, 8 tun khi trn my phn tch
Cc loi dung dch h thng khc
- Chun, nc mui sinh l
- Control: 2 mc
- Vt t tiu hao: ng ly mu, kim tim, bng, cn, gng tay
3. Ngi bnh: c gii thch trc khi thc hin XN, tt nht l nhn n sng v ly mu vo bui sng.
4. Phiu xt nghim: c y thng tin v ngi bnh bao gm tn, tui, khoa phng, chn on, tnh trng mu, tn BS ch nh, cc loi thuc s dng (nu c)
III. CC BC TIN HNH
1. Ly bnh phm: bnh phm phi c ly ng k thut vo ng tiu chun. Ly tm trc khi tin hnh k thut. C th s dng huyt thanh hoc huyt tng chng ng bng heparin. Bo qun 2-80C trong vng 5 ngy, - 200C c 6 thng. R ng mt ln.
-
23
bnh phm, chun, control nhit phng (20-250C) v lc u trc khi tin hnh XN.
2. Tin hnh k thut
- My mc, ha cht c ci t v chun trc khi thc hin phn tch. Control nm trong min cho php ty thuc vo k thut, thuc th ca tng cng ty. Thng thng chy control 2 min: bnh thng v bt thng. i chiu vi lut v ni kim cht lng nu t th tin hnh phn tch mu.
- a bnh phm vo phn tch theo protocol ca my. Khi c kt qu th phn tch v i chiu vi phiu xt nghim v tr kt qu.
IV. NHN NH KT QU
- Bnh thng:
Nam: 202 416 mol/l
N: 143 399 mol/l
- Acid uric mu tng trong:
Bnh Goutte
Suy thn
Nhim c ch, thu ngn
- cid uric mu gim trong:
Bnh Willson
Cn lit chu k
Xanthin niu
V. NHNG SAI ST V X TR.
Nguyn nhn Sai st X tr
Bnh phm ly vo ng chng ng bng EDT
Lm gim kt qu khong 7%
Khng s dng loi cht chng ng ny
Bnh phm tng bilirubin, huyt tn, tng lipid mu, ang s dng thuc
Kt qu nh hng khng r
Nng > di o (11,9 1487 mol/L)
Sai lch kt qu. Rt t gp
Pha long bnh phm
-
24
4. NH LNG ADH
(Anti Diuretic Hormone)
I. NGUYN L - nh lng ADH trong huyt tng v huyt thanh bng phng php min dch
enzyme (ELISA)
- Khi thc hin kit, cc ging c trng trc vi mt khng th c bit tit ra t hormone DH. Sau cht chun hay mu bnh phm c a vo cng vi Biotin-conjugated v HRP cho mi ging ri . Tip cht th TMB thm vo. Ch nhng ging c cha ADH, biotin-conjugated khng th v enzyme-conjugated Avidin s lm i mu. Nng ca DH c xc nh trn th v t nng mu o OD vi nng chun.
- Nng chun: 80 pg/ml, 40 pg/ml, 20 pg/ml, 10 pg/ml, 5pg/ml, 2.5 pg/ml, 1.25 pg/ml.
II. CHUN B 1. Ngi thc hin
Ngi thc hin xt nghim c trnh ph hp 2. Phng tin, ha cht 2.1 Vt liu t chun b
- Micropipettes (10-100L/100-1000L) - Ging 96 l - My Elisa - My o quang - My
2.2 Ha cht v thuc th trong b Kit ca nh cung cp - Wash Buffer (nng 25X) - Cht chun - Mu pha long - Biotin-antibody pha long - HRP-avidin pha long - Biotin-antibody - HRP-avidin - Stop Solution - Thuc th TMB iu kin bo qun : 2-8oC
3. Ngi bnh
Ngi bnh cn c t vn v mc ch lm xt nghim.
-
25
- gp phn chn on mt s tnh trng bnh l c th gy bi xut bt thng DH (V d: hi chng tit DH khng thch hp [SI DH]).
- gp phn chn on mt s tnh trng bnh l gy mt bi xut DH hay gy mt p ng thn i vi tc dng ca DH (V d: i nht ngun gc trung ng v i nht ngun gc thn).
- chn on v chn on phn bit gia i tho nht vi tnh trng i nhiu do cn nguyn thn kinh (psychogenic polyuria).
- chn on phn bit cc trng hp gim natri mu.
4. Phiu xt nghim
Phiu xt nghim cn ghi y thng tin v tn, tui, gii tnh, khoa phng, chn on ca ngi bnh v ghi r ch nh xt nghim.
III. CC BC TIN HNH 1. Ly bnh phm
Xt nghim c tin hnh trn huyt tng - Yu cu ngi bnh nhn n 10-12 gi trc khi ly mu xt nghim. Ngi
bnh c yu cu trnh cc hot ng th lc v b stress trong thi gian xt nghim. Ly mu khi ngi bnh t th ngi.
- Mu bnh phm sau khi ly cn c bo qun trong ti lnh v c chuyn ngay ti phng xt nghim (khng c trong iu kin nhit phng).
2. Tin hnh k thut ADH ELISA
- Thm 100 l cht chun, Blank, mu bnh phm vo mi ging. y np v trong 2 gi 370C
- Ht b cht lng trn b mt ging. Ch : khng ra
- Thm 100 l Biotin-antibody, 1 gi 370C
- Ht ht c cht ri ra vi 200l Wash Buffer - Lp li quy trnh 3 ln
- Thm 100 l HRP-avidin vo mi ging, 1 gi 370C
- Ra 5 ln vi 200l Wash Buffer
- Thm 90l thuc th TMB vo mi ging, 10-30pht 370C
- Thm 50l dung dch Stop Solution ngng phn ng khi 4 ging u tin cha nng cht chun cao nht i sang mu xanh. Nu s i mu khng xut hin ng b, phi chc l cc cc trong mi ging c trn u.
- c hp th ti bc sng 450nm trong 10 pht. IV. NHN NH KT QU
S dng ng cong chun tnh ton nng ADH trong mu: - To mt ng cong chun bng cch x l phn tch d liu dng phn mm
my tnh thch hp thng k 4 tham s.
-
26
- Trc y: nng mu bnh phm, trc x: nng cht chun. - Trung bnh kt qu lp li ca mi ln o cht chun, control, sample v tr cho
trung bnh mt quang ca standard zero.
Kt lun: kt qu c bin lun a ra kt lun da trn cc tr s tham chiu. V. NHNG SAI ST V X TR
- Nu gi tr nng mu bnh phm cao hn nng cht chun mc cao nht. Pha long mu vi cht Standard Diluent v lp li quy trnh.
- Khi s dng my ra t ng phi cng thm khong 30 giy sau khi thm m ra v lc 180 gia cc bc ra s lm gia tng s chnh xc.
m bo kt qu xc thc, np y cn phi dnh st trong qu trnh
-
27
5. NH LNG ADIPONECTIN
diponectin l mt polypeptid gm 244 axit amin. diponectin ch yu c tng hp bi cc t bo m. Mc diponectin thp lin quan cht ch vi s khng insulin v hi chng chuyn ha, cng nh tng nguy c i tho ng type 2 v bnh tim mch.
I. NGUYN L
Dng k thut ELIS nh lng diponectin trong nui cy t bo, huyt thanh v huyt tng ngi
Da vo tnh c hiu ca khng nguyn - khng th theo phng php sandwich: cc ging c ph khng th c hiu cho diponectin ngi. Standard, mu v Biotin-khng th c thm vo, diponectin trong mu kt hp vi khng th ph trn ging v Biotin-khng th mi c thm vo. Sau khi ra i cc Biotin-khng th khng kt hp, th HRP- Avidin lin hp c thm vo. Tip theo, sau khi cc ging c ra ln hai, c cht TMB c thm vo ging. Sau , dung dch ngng phn ng thm vo s chuyn dung dch phn ng t mu xanh sang mu vng, m mu t l thun vi nng diponectin trong mu, c o bc sng 450 nm.
II. CHUN B
1. Ngi thc hin
Bc s, c nhn, k thut vin c o to vi my Evolis Twin Plus
2. Phng tin, ha cht
- My phn tch ELIS (c th Evolis Twin Plus)
- Thuc th c cung cp ca hng DRG (c) (EI -4820)
+ a phn ng (96 ging)
+ HRP- Avidin lin hp
+ Chun: 2 l (dng ng kh)
+ C cht TMB
+ Biotin-khng th diponectin
+ Dung dch ra
+ Dung dch pha long
+ Dung dch ngng phn ng
-
28
Trong : HRP- vidin l vidin lin kt vi HRP (Horseradish Peroxidase). TMB: 3,3,5,5 tetramethyl-benzidine.
- Thuc th v dng c cn nhng khng c cung cp
+ Pipet chnh xc
+ Cc tube
+ u cn pipet dng mt ln
+ Nc ct
+ Control level 1 v 2.
3. Ngi bnh
Ngi bnh khng cn nhn i hay yu cu g.
4. Phiu xt nghim
Phiu xt nghim theo mu bnh vin v B Y t quy nh, c ghi y thng tin ngi bnh.
III. CC BC TIN HNH
1. Ly bnh phm
S dng huyt thanh, huyt tng, dung dch ni trong nui cy t bo
- Huyt tng:
+ Thu thp huyt tng bng cch s dng mt phn mi th tch natri citrat 0,1M nh mt cht chng ng. Ly tm mu vi 3000 vng trong 10 pht.
+ Pha long mu huyt tng vi dung dch pha long theo t l 1:1000. Nn thc hin pha long theo hai bc, v d:
B1: Pha long 1:100 (5 mu + 495 dung dch pha long)
B2: Pha long 1:10 (100 mu pha long + 900 dung dch pha long).
+ Khng s dng mu c, mu v hng cu.
+ Mu cha pha long c th c lu tr -200C hoc thp hn. Ch r ng mt ln
- Huyt thanh:
+ Sau khi hnh thnh cc mu ng, ly tm mu vi 3000 vng trong 10 pht.
+ Pha long mu huyt thanh vi dung dch pha long theo t l 1:1000. Nn thc hin pha long theo hai bc nh trn.
+ Mu cha pha long c th c lu tr -200C hoc thp hn, n nh c 3 thng. Ch r ng mt ln.
-
29
- Dung dch ni trong nui cy t bo
+ Ly tm vi 3000 vng/ pht, trong10 pht loi b cc mnh vn.
+ Pha long dung dch ni trong nui cy t bo vi dung dch pha long mu theo t l 1:10
(V d: 10 mu + 90 dung dch pha long)
+ Lu tr -200C hoc thp hn. Ch r ng mt ln.
2. Tin hnh k thut
2.1. Chun b thuc th
a tt c thuc th v nhit phng trc khi s dng.
2.1.1. Dung dch ra:
Ha 50 ml dung dch ra vi 950 ml nc ct c dung dch 1000 ml.
Sau khi pha, n nh 3 ngy nhit phng, 2 tun 80C
2.1.2. Chun:
- L chun gc c nng 30 ng/mL
- Pha long nhiu ln vi dung dch pha long c S4 - S1vi cc nng tng ng l 15 ng/mL; 5 ng/mL; 1,67 ng/mL; 0,56 ng/mL; 0,185 ng/mL v S0 l dung dch pha long (0 ng/mL)
- Chun th khng n nh sau khi pha long s dng ngay sau khi pha long hoc lm ng nu khng s dung trong vng 1 gi.
c. Biotin-khng th v HRP-Avidin
Votex u, nh nhng trc khi dng.
2.2. Tin hnh
- Tin hnh theo quy trnh ci t trn my t ng EVOLIS TWIN PLUS.
- Thi gian hon thnh xt nghim ny khong 255 pht
- V ng cong chun trc, control t th tin hnh o mu.
Cc bc tin hnh nh sau:
+ Ht 50 l mi calibrator/ QC hoc mu ngi bnh vo cc ging
+ 120 pht nhit phng
+ Ra cc ging 5 ln vi 300 dung dch ra cho mi ging trong mt ln ra
+ Ht 50 l Biotin - khng th vo mi ging
+ 60 pht nhit phng
-
30
+ Ra cc ging 5 ln vi 300 l dung dch ra cho mi ging trong mt ln ra
+ Ht 50 l HRP- vidin lin hp vo mi ging
+ 30 pht nhit phng
+ Ra cc ging 5 ln vi 300 l dung dch ra cho mi ging trong mt ln ra
+ Ht 50 l c cht TMB vo mi ging.
+ 20 pht nhit phng.
+ Ht 25 l dung dch ngng phn ng vo mi ging
+ Tin hnh o vi bc sng 450/630 nm ngay lp tc
IV. NHN NH KT QU
- Gi tr tham kho:
Nam trng thnh: 8000 10000 ng/mL (8 -10 mg/L)
N trng thnh : 10000 12000 ng/mL (10 - 12 mg/L)
- ngha lm sng: diponectin gim c nguy c b cc bnh sau: Bnh bo ph X va ng mch Bnh ng mch vnh Hi chng chuyn ha
B sung omega-3, tng cng cht x trong khu phn hng ngy, hoc tp th dc iu gip lm tng nng adiponectin.
V. NHNG SAI ST V X TR
C mt s sai st thng gp:
- Ly sai ng ly li
- Tuyt i khng s dng mu v hng cu, mu c, mu vng
- Mu mu ngi bnh c dng thuc chng ng th thi gian co cc mu lu hn trc khi ly tm (hn 30 pht)
- Mu c kt qu vt qu 15 ng/mL th phi ha long mu vi dung dch pha long.
- Nhng sai st do my th hi k s x tr.
- Lu Calibrator v QC bo qun tht tt c ng cong chun t yu cu.
-
31
6.NH LNG ALDOSTEROL
ldosteron l hormon tuyn v thng thn tng s ti hp thu Natri ng thn dn ti vic bi tit Kali, iu chnh lng mu tun hon. Sn sinh tha v bi tit thng xuyn (mn) ldosteron s dn ti chng tng huyt p. o mc ldosteron trong huyt thanh cng vi rennin huyt thanh c th c dng phn bit gia ldosteronism tin pht v th pht.
I. NGUYN L - Aldosteron c nh lng bng k thut ELISA . - Mu bnh phm (huyt thanh) c thm vo cc ging c trng bng khng th
khng Aldosteron v trong dung dch m 30 pht nhit phng. Nu Aldosteron trong mu bnh phm, s kt hp vi cc khng th c trong ging. Ging c ra loi b mu d v khng th khng Aldosteron gn peroxidase c thm vo s to phc lin kt enzyme v khng th kp gia l phn t Aldosteron.
- Sau khi nhit phng 30 pht, cc ging c ra loi b cc thnh phn d tha. C cht TMB c thm vo tip dung dch phn ng nhit phng trong 20 pht chuyn thnh mu xanh da tri. Tin hnh c phn ng bc sng 450 nm.
- Nng Aldosteron t l vi m ca mu xanh trong ging II. CHUN B 1. Ngi thc hin
Ngi thc hin xt nghim c trnh ph hp. 2. Phng tin, ha cht
- Pipettes th tch 50, 100, 150 v 300 microL - u pipet dng 1 ln - Nc ct hoc nc kh Ion - 3.2 N HCL v 3,2 N NaOH (dng cho xt nghim nc tiu) - ng thy tinh hay Polypropylene (dng cho xt nghim nc tiu) - Nc ra - My lc a - My c a ging vi knh lc 450nm v OD > 3.0 hoc cao hn
2.1. Cc cht c cung cp - a cha Khng th khng-Aldosteron t th:
+ Thnh phn: 1 da cha 96 (12x8) ging trong ti np ko, c cha cht ht m + Lu tr: 2-8C + n nh: 12 thng hoc xem hng dn trn bao b
- Aldosterone-Biotin: Avidin-Horse Radish Peroxidase (HRP) Conjugate Concentrate X50:
-
32
+ Thnh phn: Aldosterone-Biotin v Avidin-Horse Radish Peroxidase (HRP) Conjugate Concentrate trong dung dch m gc protein khng cha thy ngn
+ Th tch: 300 microL/l + Lu tr: 2-8C + n nh: 12 thng hoc xem hng dn trn bao b + Chun b: Pha long Aldosterone-Biotin v Avidin-HRP Concentrate 1:50 trong
dung dch m xt nghim trc khi dng. Nu dng ht a th pha 240 microL HRP trong 12 ml dung dch m xt nghim. b phn tha nu c
- Aldosteron Cht chun-C sn:
+ Thnh phn: 6 l cha ldosterone trong dung dch m gc protein khng cha thy ngn
Calibrator Concentration Volume/Vial
Calibrator A 0 pg/ml 2.0 ml
Calibrator B 20 pg/ml 0.5 ml
Calibrator C 80 pg/ml 0.5 ml
Calibrator D 300 pg/ml 0.5 ml
Calibrator E 800 pg/ml 0.5 ml
Calibrator F 2000 pg/ml 0.5 ml
+ Lu tr: 2 8C + n nh: 12 thng hoc xem hng dn trn bao b. Khi m ra th phi dng
trong vng 14 ngy hoc lu tr ng lnh. Trnh ng v r ng nhiu ln - Cht chng: C sn:
+ Thnh phn: 02 l cha Aldosterone trong dung dch m gc protein khng cha thy ngn. Tham kho nhn l bit gii hn s dng
+ Th tch: 0.5 mL/l + Lu tr: 2-80C + n nh: 12 thng hoc xem hng dn trn bao b. Khi m ra th phi dng
trong vng 14 ngy hoc lu tr ng lnh. Trnh ng v r ng nhiu ln - Wash Buffer Concentrate X10:
+ Thnh phn: 01 chai cha dung dch m khng thy ngn v cha cht ty non-ionic.
+ Th tch: 50 mL/chai + Lu tr: 2-80C + n nh: 12 thng hoc xem hng dn trn bao b. + Chun b: Pha long 1:10 trong nc ct hoc nc kh Ion. Nu dng ht a th pha 50ml trong 450ml nc
- Assay Buffer C sn:
-
33
+ Thnh phn: 01 l cha dung dch m gc protein khng thy ngn. + Th tch: 15 ml/l + Lu tr: 2-80C + n nh: 12 thng hoc xem hng dn trn bao b.
- m TMB C sn: + Thnh phn: 01 chai cha Tetramethylbenzidine v Hydrogen Peroxide trong
Non-DMF hoc DMSO cha dd m. + Th tch: 16 ml/chai + Lu tr: 2-80C + n nh: 12 thng hoc xem hng dn trn bao b.
- Dung dch ngng C sn: + Thnh phn: 01 chai cha 1M Slfuric Acid. + Th tch: 6 ml/chai + Lu tr: 2-80C + n nh: 12 thng hoc xem hng dn trn bao b.
3. Ngi bnh
Cn c t vn v mc ch xt nghim:
- chn on tnh trng cng Aldosteron tin pht hay th pht. - nh gi sn xut Aldosteron thng thn. - chn on phn bit cc ri lon nc v in gii. 4.Phiu xt nghim
Phiu xt nghim cn ghi y thng tin v tn, tui, gii tnh, khoa phng, chn on ca ngi bnh v ghi r ch nh xt nghim.
III. CC BC TIN HNH
- X l mu: - Huyt thanh: khng cn - Nc tiu: Thy phn, Trung ha v Pha long
- Tt c cc cht cn phi t nhit phng trc khi dng. Mu bnh, mu chng, mu chun phi c xt nghim 2 ln. Khi mt quy trnh bt u th phi thc hin cc bc hon chnh, khng c ngng li. - Chun b dung dch conjugate v dung dch ra - Tho nhng di s trn a vi ging. Dn ming ti v ct nhng phn cha dng vo t ng
- Dng pipet ht 50 l mi mu bnh (huyt thanh hoc nc tiu), mu chun, mu chng a vo ging tng ng, lm 2 ln.
- Dng pipet ht 100 l dung dch conjugate cho vo mi ging (nn dng pipet loi a knh)
- trn my lc a (200 vng/pht) trong 1 gi nhit phng.
-
34
- Ra ging 3 ln bng 300 l nc ra m pha long cho mi ging v np cht a li bng giy ht m bo m a kh
- Dng pipet ht 150 l cht nn TMB cho vo mi ging cng mt khong thi gian nh nhau
- trn my lc a t 10-15 pht nhit phng (hoc cho ti khi Calibraror t mu xanh bin m)
- Ht 50 l dd dng cho mi ging cng khong thi gian nh trong bc 7 - c a trn my c 450 nm trong 20 pht sau khi thm dung dch ngng vo (Trnh t v thi gian thc hin xt nghim theo b kit ca cng ty DRG Internaltional, Inc., M)
IV. NHN NH KT QU
- Tnh ton mt quang ca mi mu chun lm 2 ln - V ng chun trn giy semi-log vi trc Y l mt quang v trc X l nng cht chun. - Tnh ton mt quang ca mu - c gi tr ca mu huyt thanh trc tip t ng cong chun. Nu kt qu cao hn 2000pg/ml th phi pha long cht chun t l khng qu 8 ln, khi gi tr o c phi nhn vi h s pha long. - c gi tr ca mu nc tiu trc tip t ng cong chun v nhn vi h s 60 t. K tip, nhn vi lng nc tiu trong vng 24 gi thu thp c (lt). Cui cng chia vi 1000 t c gi tr g/24 gi. Nu kt qu ln hn 2000 pg/ml, pha long cht chun nhng khng qu 2 ln, kt qu nhn li vi h s pha long. - Nng ldosteron thay i trong cc trng hp:
Bnh ldosteron huyt thanh Renin huyt tng
Aldosteronism tin pht Cao Thp
ldosteronism th pht Cao Cao
V. NHNG SAI ST V X TR
1. Lu v cnh bo
- Mu kim tra c mc cao v thp v bnh phm huyt thanh phi cng tin hnh mi ln nh gi tin cy ca kt qu.
- Phi dng nc ct hoc nc kh Ion pha long - Phi eo gng tay khi lm th nghim - Tt c mu phi c n nh nhit phng trc khi dng. Trnh vic ng v tan ng mu lp li
- ng chun phi c thit lp cho mi ln chy
-
35
- Trnh bt kh trong ging (Microwell) s nh hng ti mt quang hc (ODs). Cn thn loi b bt kh trc khi c mu.
- Dung dch nn (TMB) rt nhy sng v phi duy tr trong khi lu tr thch hp. Khi dung dch c mu xanh th c ngha l dung dch b nhim bn, khng n nh v s khng c dung.
- Khi pha ch dung dch nn v dung dch dng phn ng, khng c dng pipet do nguy c tip xc vi kim loi
- Cc cht v dung dch tha sau th nghim phi c xem nh l cht thi c hi v phi tiu hy theo quy nh.
2. Hn ch
- Tt c cc tc cht c cn chnh dng xc nh trc tip ldosteron trong huyt thanh v nc tiu ngi. B kit ny khng dng cho cc mu vt khc ca ngi hoc ng vt
- Khng dng huyt thanh bo qun khng ng, huyt thanh b tn huyt v cc vn khc
- Bt k mu hay huyt thanh chng cha azide hoc thimerosal u khng tng thch vi kit ny
- Ch c Cht chun mi c th dng pha long mu huyt thanh hoc nc tiu. Dng cc tc cht khc c th lm kt qu sai lch.
-
36
7. NH LNG ALBUMIN
I. NGUYN L
nh lng lbumin trong mu ca ngi bnh theo phng php so mu
pH= 4.1
Albumin + BCG => Albumin BCG complex
(BCG: Bromcresol green)
Phc hp lbumin BCG c mu xanh t l thun vi nng lbumin trong mu th c o bc sng 570 nm.
II. CHUN B
1. Ngi thc hin
01 cn b i hc, 1 k thut vin chuyn ngnh ha sinh
2. Phng tin, ha cht
- Phng tin: My xt nghim nh Cobas C501, U 640.
- Ha cht: Ha cht xt nghim lbumin, cht chun lbumin, cht kim tra cht lng lbumin.
3. Ngi bnh
Ngi bnh cn c gii thch v mc ch ca vic ly mu lm xt nghim.
4. Phiu xt nghim
Phiu xt nghim cn ghi y thng tin v tn, tui, gii tnh, khoa phng, chn on ca ngi bnh v ghi r ch nh xt nghim
III. CC BC TIN HNH
1. Ly bnh phm
- Ly 3 ml mu tnh mch vo ng khng c cht chng ng hay ng c cht chng ng l Heparin, EDT , khng s dng cht chng ng Fluorid. Mu khng v hng cu. Bnh phm n nh 5 thng 28C, 2.5 thng 15 - 25C. 4 thng -15 n -25C.
- Sau khi ly mu, em ly tm tch ly huyt thanh hoc huyt tng.
- Bnh phm ch r ng 1 ln v phi bnh phm t nhit phng trc khi phn tch. trnh hin tng bay hi, bnh phm, cht chun, cht kim tra cht lng nn phn tch trong vng 2 gi.
-
37
2. Tin hnh k thut
- My phn tch cn chun b sn sng thc hin phn tch mu: My c ci t chng trnh xt nghim lbumin. My c chun vi xt nghim lbumin. Kt qu kim tra cht lng vi xt nghim lbumin t yu cu khng nm ngoi di cho php v khng vi phm lut kim tra cht lng.
- Ngi thc hin phn tch mu nhp d liu v thng tin ngi bnh v ch nh xt nghim vo my phn tch hoc h thng mng (nu c).
- Np mu bnh phm vo my phn tch
- Ra lnh cho my thc hin phn tch mu bnh phm
- i my phn tch mu theo protocol ca my
- Khi c kt qu cn xem xt nh gi kt qu sau in bo co hoc ghi kt qu vo phiu xt nghim tr cho ngi bnh.
IV. NHN NH KT QU
- Tr s bnh thng: 34 48 g/l.
- lbumin mu tng trong: Mt nc (nn nhiu, tiu chy nng).
- Albumin mu gim trong: Bnh thn (suy thn, hi chng thn h, vim cu thn). Bnh khng c albumin huyt bm sinh. Gim tng hp (vim gan nng, x gan), km hp thu, km dinh dng, Mt albumin (bng, tn thng r dch, bnh ng rut mt protein). Ung th, nhim trng.
V. NHNG SAI ST V X TR
Nhng yu t gy nhiu cho kt qu xt nghim. Kt qu xt nghim khng b nh hng khi:
- Huyt thanh vng: Bilirubin < 60 mg/dL hay 1026 mol/L.
- Tn huyt: Hemoglobin < 1000 mg/dL.
- Huyt thanh c: Triglyceride
-
38
8. NH LNG ALPHA 1 ANTITRYPSIN
Alpha-1 antitrypsin, cn c gi l AAT, l mt protein do gan sn xut, lu thng trong mu. N gip bo v cc c quan ca c th khi nhng tc ng c hi ca cc protein khc. Thiu AAT l mt bnh c tnh di truyn. Nhng c th b thiu ht AAT c th lin quan n bnh l v phi hoc bnh gan (bnh hen, kh ph thng; x gan)
I . NGUYN L
nh lng T bng phng php min dch o c. Bnh phm c cho thm thuc th 1, sau cho thm thuc th 2 (c cha khng th khng lpha1 ntitrypsin. Lc ny xy ra phn ng kt hp gia khng th v khng nguyn c trong mu bnh phm, to thnh phc hp ngng kt. C th xc nh c c t xc nh c nng lpha1 ntitrypsin thng qua ng chun.
II. CHUN B
1. Ngi thc hin: 01 cn b i hc chuyn ngnh Ha sinh v 01 k thut vin
2. Phng tin
2.1. Phng tin
- Cc my c th phn tch: modular analytics e170, cobas 6000, cobas 8000, AU 640, 680, 2700, 5800 v mt s my khc.
- My ly tm
- T lnh bo qun ha cht v bo qun QC, mu bnh phm
- Pipet cc loi, ng sample cup
- ng nghim, u cn xanh v vng
- Gi ng ng nghim
2.2. Ha cht
- Thuc th 1 (R1): m Phosphate: 12.7 mmol/L, pH 7.2; NaCl: 0.13 mol/L; PEG: 40 g/L; cht bo qun
- Thuc th 2 (R2) Anti-human 1-antitrypsin antibody: > 2 g/L; NaCl: 0.12 mol/L; cht bo qun
- Dung dch chun (Cht chun)
- Dung dch QC (cc mc )
2.3. Cc dng c tiu hao khc
- ng nghim
-
39
- Gng tay
- Bng , cn st trng, bm tim hoc kim ly mu
3. Ngi bnh
Cn gii thch cho ngi bnh v ngi nh v mc ch v ngha ca xt nghim
Ngi bnh cn phi hp ly mu theo ng yu cu v thi gian v s lng
4. Phiu xt nghim
Thc hin theo y lnh ca bc s lm sng trn phiu xt nghim
III. CC BC TIN HNH
1. Ly bnh phm
C th s dng Huyt thanh, hoc Huyt tng: cht chng ng Li-heparin, EDTA v citrate
Mu c th n nh:
7 ngy / nhit 20-25C
3 thng / nhit - 20C, ch c php ng mt ln, mu c vn ta cn ly tm trc khi phn tch
2. Tin hnh k thut
2.1. Chun b my phn tch
Dng ng chun
Phn tch QC: c 2 level. Khi QC t tin hnh phn tch mu
2.2. Phn tch mu
Mu bnh phm nn c tin hnh phn tch trong vng 2h
Mu sau khi ly tm c chuyn vo khay ng bnh phm
nh s (hoc ID ca ngi bnh); chn test v my s t ng
IV. NHN NH KT QU
Tr s tham kho: 16,6 36,8 mol/L (90 200 mg/dL hoc 0,9 2,0 g/L)
H s chuyn i:
mg/dL x 0,184 = mol/L
mg/dL x 0,01 = g/L
g/L x 100 = mg/dL
-
40
V. NHNG SAI ST V X TR
Cc yu t c th gy nh hng n kt qu xt nghim khi:
Hemoglobin > 621 mol/L (1000 mg/dL)
Yu t dng thp > 100 IU/mL
X tr: khi ly mu trnh gy v hng cu. Khi chun b thy mu mu b v hng cu nn loi v ly mu mu khc.
-
41
9. O HOT PHOSPHATASE KIM
(Alkaline phosphatase - ALP)
Phosphatase kim l mt enzym gan c bi tit theo dch mt, thng tng khi c tc mt. Ngoi ra mt s cc nguyn nhn khc cng c th tng phosphatase kim nh bnh xng...
I. NGUYN L
Hot ca enzym LP trong mu ca ngi bnh c xc nh theo phng php ng hc enzym.
ALP
p-nitrophenyl phosphate + H2O => phosphate + p-nitrophenol
Mg2+
p-nitrophenol c to thnh t l thun vi hot LP v c o bc sng 405 nm.
II. CHUN B
1. Ngi thc hin
01 cn b i hc, 01 k thut vin chuyn ngnh ha sinh
2. Phng tin, ha cht
- Phng tin: My xt nghim t ng nh Cobas C501, U 640.
- Ha cht: Ha cht xt nghim LP, cht chun LP, cht kim tra cht lng LP.
3. Ngi bnh
Ngi bnh cn c gii thch v mc ch ca vic ly mu lm xt nghim.
4. Phiu xt nghim
Phiu xt nghim cn ghi y thng tin v tn, tui, gii tnh, khoa phng, chn on ca ngi bnh v ghi r ch nh xt nghim
III. CC BC TIN HNH
1. Ly bnh phm
- Ly 3 ml mu tnh mch vo ng khng c cht chng ng hay ng c cht chng ng l Li, Na hay NH4-heparin. Mu khng v hng cu. Bnh phm n nh 7 ngy 28C, 2 thng -15C n -25C .
-
42
- Sau khi ly mu, em ly tm tch ly huyt thanh hoc huyt tng.
- Bnh phm ch r ng 1 ln v phi bnh phm t nhit phng trc khi phn tch. trnh hin tng bay hi, bnh phm, cht chun, cht kim tra cht lng nn phn tch trong vng 2 gi.
2. Tin hnh k thut
- My phn tch cn chun b sn sng thc hin phn tch mu: My c ci t chng trnh xt nghim LP. My c chun vi xt nghim LP. Kt qu kim tra cht lng vi xt nghim LP t yu cu khng nm ngoi di cho php v khng vi phm lut kim tra cht lng.
- Ngi thc hin phn tch mu nhp d liu v thng tin ngi bnh v ch nh xt nghim vo my phn tch hoc h thng mng (nu c).
- Np mu bnh phm vo my phn tch
- Ra lnh cho my thc hin phn tch mu bnh phm
- i my phn tch mu theo protocol ca my
- Khi c kt qu cn xem xt nh gi kt qu sau in bo co hoc ghi kt qu vo phiu xt nghim tr cho ngi bnh.
IV. NHN NH KT QU
+ Tr s bnh thng:
Nam: 40 129 U/L.
N: 35 104 U/L.
+ LP mu tng trong: Cng cn gip, Thiu Vitamin D, Bnh xng (ci, mm, x cng, ung th, sarcom). Bnh gan (Tc mt ngoi gan, Vim ng mt, K gan, bces gan), bnh thn
+ LP mu gim trong: gim LP gia nh, suy gip, thiu Vitamin C
V. NHNG SAI ST V X TR
+ Nhng yu t gy nhiu cho kt qu xt nghim
Kt qu xt nghim khng b nh hng khi:
- Huyt thanh vng: Bilirubin < 70 mg/dL hay 1197 mol/L.
- Tn huyt: Hemoglobin < 500 mg/dL.
- Huyt thanh c: Triglyceride
-
43
10. O HOT AMYLASE
I. NGUYN L
mylase l enzyme thy phn tinh bt, c ngun gc t ty v tuyn nc bt. Xt nghim amylase thng c ch nh trong bnh l tuyn ty hoc tuyn nc bt
Hot ca enzym mylase trong mu ca ngi bnh c xc nh theo phng php ng hc enzym.
-amylase
5 ethylidene-G7PNP + 5 H2O 2 ethylidene-G5 + 2 G2PNP + 2 ethylidene-
G4 + 2 G3PNP +ethylidene-G3 + G4PNP
-glucosidase
2 G2PNP + 2 G3PNP + G4PNP + 14 H2O 5 PNP + 14 G
m mu sc ca PNP hnh thnh t l thun vi hot amylase huyt thanh v c th o c bc sng 415 nm
II. CHUN B
1. Ngi thc hin
01 i hc, 01 k thut vin chuyn ngnh ha sinh.
2. Phng tin, ha cht
- Phng tin: My xt nghim nh Cobas C501, U 640, MODUL R P, Hitachi 902, 912.
- Ha cht: Ha cht xt nghim mylase, cht chun mylase, cht kim tra cht lng Amylase.
3. Ngi bnh
Ngi bnh cn c gii thch v mc ch ca vic ly mu lm xt nghim.
4. Phiu xt nghim
Phiu xt nghim cn ghi y thng tin v tn, tui, gii tnh, khoa phng, chn on ca ngi bnh v ghi r ch nh xt nghim.
III. CC BC TIN HNH
1. Ly bnh phm
-
44
- Ly 3 ml mu tnh mch vo ng khng c cht chng ng hay ng c cht chng ng l Li, Na hay NH4-heparin hoc EDT (nu dng EDT , kt qu thp hn 5-10% so vi huyt thanh). Mu khng v hng cu. Bnh phm n nh 1 thng 28C, 7 ngy 20C n 25C .
- Sau khi ly mu, em ly tm tch ly huyt thanh hoc huyt tng.
- Bnh phm ch r ng 1 ln v phi bnh phm t nhit phng trc khi phn tch. trnh hin tng bay hi, bnh phm, cht chun, cht kim tra cht lng nn phn tch trong vng 2 gi.
2. Tin hnh k thut
- My phn tch cn chun b sn sng thc hin phn tch mu: My c ci t chng trnh xt nghim mylase. My c chun vi xt nghim mylase. Kt qu kim tra cht lng vi xt nghim mylase t yu cu khng nm ngoi di cho php v khng vi phm lut kim tra cht lng.
- Ngi thc hin phn tch mu nhp d liu v thng tin ngi bnh v ch nh xt nghim vo my phn tch hoc h thng mng (nu c).
- Np mu bnh phm vo my phn tch
- Ra lnh cho my thc hin phn tch mu bnh phm
- i my phn tch mu theo protocol ca my
- Khi c kt qu cn xem xt nh gi kt qu sau in bo co hoc ghi kt qu vo phiu xt nghim tr cho ngi bnh.
IV. NHN NH KT QU
+ Tr s bnh thng: < 100 U/L
+ mylase mu tng trong: Bnh tu (vim tu cp v mn), Bnh ng mt, Bnh bng khng phi bnh tu (lot thng d dy, tc rut...), Quai b, tc tuyn nc bt, Tng mylase ngi bnh thng (tng Macro mylase)
+ mylase gim khi ty b hoi t lan rng, ngoi ra n cn gim trong mt s bnh l nh: Vim tu mn tnh. Vim ty mn tnh tin trin. X ho ng dn ty tin trin.
V. NHNG SAI ST V X TR
Nhng yu t gy nhiu cho kt qu xt nghim. Kt qu xt nghim khng b nh hng khi:
- Huyt thanh vng: Bilirubin < 60 mg/dL hay 1026 mol/L.
- Tn huyt: Hemoglobin < 500 mg/dL.
- Huyt thanh c: Triglyceride
-
45
Khc phc: C th ha long bnh phm v thc hin li xt nghim sau nhn kt qu vi ha long (Trng hp c ha long t ng trn my th kt qu khng cn nhn vi ha long do my t tnh ton kt qu)
Lu : Nc bt v m hi c cha mylase nn trnh nhim nhng cht ny vo bnh phm hay ha cht.
-
46
11. NH LNG AMONIAC
Trong iu kin bnh thng moniac c chuyn i thnh ure ti gan v sau c thn bi xut. Nu c mt ri lon thc th ngn khng cho qu trnh chuyn i ni trn xy ra, NH3 s tch t trong dng tun hon. Khi tch t trong mu vi nng cao s gy ra mt tnh trng bnh no gan.
I . NGUYN L
Theo phng php ng ng hc enzym. Da trn phn ng sau:
Enzym glutamate dehydrogenase (GLDH) xc tc phn ng vi s tham gia ca NADPH theo s sau:
Lng N DPH2 b oxy ha trong giai on phn ng s tng ng vi lng NH3 c trong mu bnh phm. C th o c s gim mt quang hc do NADPH chuyn thnh N DP bc sng vng t ngoi
II. CHUN B
1. Ngi thc hin
i hc chuyn ngnh Ha sinh min dch v 01 k thut vin.
2. Phng tin, ha cht
2.1. Phng tin
My phn tch ha sinh t ng: Hitachi 912, 917, cobas 6000; cc my U 640, 2700, 5800 v mt s my khc.
2.2. Ha cht
R1 Triethanolamine buffer: 151 mmol/L, pH 8.6;
-ketoglutarate: 16.6 mmol/L; DP: 1.2 mmol/L; cht bo qun
R2 N DPH: 458 mol/L; GLDH: 24.3 U/mL (EC 1.4.1.3; bovine liver; 25C); triethanolamine buffer: 151 mmol/L, pH 8.6;
-ketoglutarate: 16.6 mmol/L; DP: 1.2 mmol/L; cht bo qun.
(tham kho theo ho cht ca Roche diagnostic)
2.3. Cc dng c tiu hao khc
- ng nghim
-
47
- Gng tay, dy gar
- Bng , cn st trng, bm tim hoc kim ly mu
3. Ngi bnh
- Cn gii thch cho ngi bnh v ngi nh mc ch xt nghim
- Ngi bnh cn nhn n 8 - 10h trc khi ly mu
- Trnh hot ng th lc qa mc v ht thuc trc khi ly mu xt nghim
4. Phiu xt nghim
C y lnh ca bc s lm sng ghi trn phiu xt nghim
III. CC BC TIN HNH
1. Ly bnh phm
- Ly mu vo ng chng ng EDT
- Mu cn c y np cht, bo qun lnh v chuyn ngay xung phng xt nghim. Ti phng xt nghim cn ly tm phn tch mu cng sm cng tt.
2. Tin hnh k thut
2.1. Chun b my phn tch
Dng ng chun: da trn 2 im
Phn tch QC: c 2 level. Khi QC t tin hnh phn tch mu
2.2. Phn tch mu
Mu bnh phm nn c tin hnh phn tch trong vng 2gi.
Mu sau khi ly tm c chuyn vo khay ng bnh phm.
nh s (hoc ID ca ngi bnh); chn test v vn hnh theo protocol ca my, my s t ng phn tch.
IV. NHN NH KT QU
Tr s tham kho:
- Nam: 14.7 - 55.3 mol/L (25-94 g/dL)
- N: 11.2 - 48.2 mol/L (19-82 g/dL)
- Tr em: 28 57 mol/L (40 80 mg/L)
- Tr s sinh: 64 1072 mol/L (90 150 mg/L)
H s chuyn i: g/dL x 0.587 = mol/L
+ Tng NH3 mu: X gan, hi chng tng nit mu, Xut huyt tiu ha, Suy tim,
-
48
Suy gan, Leucemie, Bnh l tan mu tr s sinh, Vim mng ngoi tim
+ Gim NH3 mu: Tng huyt p v cn; Tng huyt p c tnh
V. NHNG SAI ST V X TR
1. Cc yu t nh hng
- Ht thuc l; Hot ng th lc vi cng cao
- Ch n c nhiu hoc qu t m (proein)
- Bilirubin lin hp > 1026 mol/L(>60 mg/dL).
- Hin tng huyt tn: khi Hb > 31 mol/L(>50 mg/dL).
- Cc thuc lm tng NH3: Heparin, thuc li tiu (furosemid)
- Cc thuc lm gim NH3: Neomyein, tetracyclin, diphenyl hydramin
2. X tr
Nhc ngi bnh trnh hot ng th lc cng cao trc thi gian ly mu.
Khi ly mu trnh gy v hng cu. Mu v hng cu cn loi v ly li mu mu khc.
-
49
12. NH LNG AMH GEN II
(Anti Mullerian Hormon)
MH l mt dimer glycoprotein gm 2 monome c trng lng phn t 72 kDa lin kt vi nhau bng cu ni disunfit. MH l hormon c ch ng Muller (hai ng phi s pht trin thnh ng sinh dc n). nam gii, MH c sn sinh bi t bo Sertoli (trong tinh hon). MH tip tc c hai tinh hon tit ra ti thi k dy th v sau gim chm dn sau dy th n mc thp (dng lng cn). n gii, MH c cc t bo ht bung trng tit ra vi mc thp sau sinh ti thi k mn kinh v sau gim ti mc khng th pht hin c.
I. NGUYN L
Dng k thut ELIS nh lng MH trong huyt thanh v huyt tng chng ng bng lithium heparin ngi.
Da vo tnh c hiu ca khng nguyn-khng th, theo phng php sandwich: Trong xt nghim ny calibrator, control v mu c trong cc ging c ph khng th MH. Sau ln v ra th nht, b sung phc hp khng th AMH Gen II-biotin vo tng ging. Sau ln v ra th hai, b sung enzym lin hp-Streptavidin vo ging. Sau ln v ra th ba, b sung c cht TMB vo ging. Cui cng b sung dung dch ngng phn ng. m mu t l thun vi nng MH c trong mu th, c o vi bc sng kp 450nm v gia 600 - 630 nm.
II. CHUN B
1. Ngi thc hin
Bc s, c nhn, k thut vin c o to vi my Evolis Twin Plus
2. Phng tin, ha cht
- My phn tch ELIS (c th Evolis Twin Plus)
- Thuc th c cung cp ca hng Beckman Coulter (M: 79765)
+ a phn ng (96 ging)
+ Phc hp khng th MH th h th 2-biotin
+ C cht TMB
+ Dung dch ra I
+ Dung dch pha long
+ Dung dch ngng phn ng A
+ Phc Enzym lin hp-Streptavidin (HRP)
-
50
+ Dung dch m
Trong : Enzym lin hp-Streptavidin l Streptavidin lin hp vi HRP (Horseradish Peroxidase). TMB: 3,3,5,5 tetramethyl-benzidine.
- Thuc th v dng c cn nhng khng c cung cp
+ AMH Gen II Calibrator v Control A79766
+ Pipet chnh xc loi 10-1000 l
+ Cc tube
+ u cn pipet dng mt ln
+ My lc, tc 600-800 vng/pht
+ My trn kiu votex
+ Vi thm ht
+ Nc kh ion
3. Ngi bnh
Khng cn nhn n v khng ph thuc chu k kinh nguyt (n)
3.1. i vi ph n: xt nghim AMH c ch nh :
- nh gi kh nng sinh sn ca bung trng
- nh gi p ng ca bung trng i vi liu php kch bung trng
- Gp phn chn on hi chng bung trng a nang.
- nh gi v theo di ung th bung trng.
- D bo thi gian mn kinh.
3.2. i vi b trai: xt nghim MH c ch nh :
- Chn on phn bit ri lon gii tnh
- Chn on tinh hon lc ch
4. Phiu xt nghim
Phiu xt nghim theo mu bnh vin v B Y t quy nh, c ghi y thng tin ngi bnh.
III. CC BC TIN HNH
1. Ly bnh phm
- S dng huyt thanh hoc huyt tng chng ng bng heparin lithium.
-
51
- co cc mu ri ly tm, tch huyt thanh ra tube, y cht np, c th bo qun 2 - 8C trong 48 gi, lu hn cn bo qun - 200C. Trnh lm ng v r ng nhiu ln.
2. Tin hnh k thut
2.1 Chun b mu
- i vi mu tr s sinh nam cn pha long mu vi dung dch pha long theo t l 1:10 trc khi xt nghim
- i vi cc mu khc th khng cn pha long.
2.2. Chun b thuc th:
a tt c thuc th v nhit phng trc khi s dng.
2.1.1.Dung dch ra:
Ha long 10 ln dung dch ra I vi nc kh ion, n nh 1 thng nhit phng.
2.1.2. Ging:
Chn s ging theo yu cu, nhng ging cn li trong bao kn c ht chng m.
2.1.3. Chun:
- T n G vi nng ln lt l: 0 ng/mL; 0,16 ng/mL; 0,4 ng/mL; 1,2 ng/mL; 4,0 ng/mL; 10,0 ng/mL v 22,5 ng/mL.
- Bo qun - 20o C sau khi nhn hng. s dng nhiu ln, chia nh v - 20oC.
- Sau ln s dng u tin, ng r ng n nh 7 ngy khi bo qun 2 8o C
- Khng r ng qu 2 ln
2.1.4. Cht chng:
- Mc 1 v mc 2
- c gi ng sau khi nhn hng - 20o C. s dng nhiu ln, chia nh v -20o C.
- Sau ln s dng u tin, ng r ng n nh 7 ngy khi bo qun 2 8o C
- Khng r ng qu 2 ln
2.1.5. Tin hnh
- Tin hnh theo quy trnh ci t trn my t ng EVOLIS TWIN PLUS.
- Tng thi gian hon thnh xt nghim ny khong 190 pht
- Tin hnh v ng cong chun, khi QC t mi tin hnh o mu
-
52
Cc bc tin hnh nh sau:
+ nh du cc ging cn s dng
+ Ht 20 l mi calibrator, control hoc mu ngi bnh vo cc ging
+ B sung thm 100 l dung dch m vo mi ging
+ trn my lc (tc khong 600 - 800 vng/pht) trong 60 pht nhit phng
+ Ra cc ging 5 ln vi 400 l dung dch ra cho mi ging trong mt ln ra
+ Ht 100 l phc hp khng th MH Gen II-biotin vo mi ging
+ trn my lc (tc 600 - 800 vng/pht) trong 60 pht nhit phng
+ Ra cc ging 5 ln vi 400 l dung dch ra cho mi ging trong mt ln ra
+ Ht 100 l phc enzym lin hp - streptavidin vo mi ging
+ trn my lc (tc khong 600-800 vng/pht) trong 30 pht nhit phng
+ Ra cc ging 5 ln vi 400 l dung dch ra cho mi ging trong mt ln ra
+ Ht 100 l dung dch to mu TMB vo mi ging. Trnh tip xc trc tip nh sng.
+ trn my lc 8-12 pht nhit phng.
+ Ht 100 l dung dch ngng phn ng vo mi ging
+ Tin hnh o
IV. NHN NH KT QU
1. i vi ph n
- nh gi kh nng sinh sn ca bung trng: Nng MH trong mu (tnh bng ng/mL) tng quan trc tip vi s lng nang trng, t thay i trong chu k kinh nguyt nn c th s dng nh gi kh nng sinh sn ca bung trng tt hn so vi FSH
- Kh nng sinh sn ti u: 4,0 - 6,8 ng/mL; kh nng sinh sn tt: 2,2 4,0 ng/mL; kh nng sinh sn km: 0,3 2,2 ng/mL; kh nng sinh sn rt km: 0,0 0,3ng/mL
- nh gi p ng ca bung trng: i vi liu php kch bung trng: Nng MH cng tng quan cht ch vi s lng non nn c th s dng nh gi p ng ca bung trng i vi liu php kch bung trng phc v cho liu php th tinh trong ng nghim IVF (In vitro fertilization) hoc liu php tim tinh trng vo bo tng ca trng ICSI (Intracytoplasmic sperm injection) trong th tinh nhn to.
-
53
- Chn on Hi chng bung trng a nang: thng MH > 6,8 ng/mL.
- nh gi v theo di ung th bung trng: nng MH c th c s dng chn on ung th t bo granulosa bung trng vi nhy khong 76 n 93%. Nng MH gim vi ngy sau phu thut khi u bung trng v li tng ln nu u ti pht
- D bo thi gian mn kinh: ph n, nng MH gim dn theo tui. MH c gi tr d bo thi k mn kinh tt hn so vi FSH. Nu MH < 0,2 ng/mL, la tui 35-39 s mn kinh sau 9,94 nm, la tui 45 48 s mn kinh sau 5,99 nm
2. i vi b trai
- Chn on phn bit ri lon gii tnh: nng MH trong mu bng 0 hoc rt thp v nng cc androgen trong mu thp: khng c tinh hon hoc khng pht trin gii tnh nam.
- Chn on tinh hon lc ch: khm khng thy c tinh hon nhng nng MH v cc androgen bnh thng
V. NHNG SAI ST V X TR
C mt s sai st thng gp:
- Ly sai ng ly li
- Tuyt i khng s dng mu v hng cu, mu c, mu vng
- Mu mu ngi bnh c dng thuc chng ng th thi gian co cc mu lu hn trc khi ly tm (hn 30 pht)
- Mu c kt qu vt qu 22,5 ng/mL th phi ha long mu vi dung dch pha long.
- Nhng sai st do my th hi k s x tr.
- Lu Calibrator v QC bo qun tht tt c ng cong chun t yu cu.
13. NH LNG ANTI-CCP
( Antibody to cyclic citrullinative peptid)
Anti CCP (Anti cyclic citrullinated peptides) l khng th khng Peptid vng cha cid min Citrulline. S hin din ca khng ny c th gip chn on bnh vim khp dng thp nhiu nm trc khi cc triu chng ca bnh xut hin.
-
54
I. NGUYN L
Anti-CCP c nh lng bng phng php min dch sandwich s dng cng ngh ha pht quang hay in ha pht quang. nti-CCP c trong mu th ng vai tr khng nguyn c kp gia hai khng th, khng th th nht l khng th n dng c hiu khng nti-CCP nh du biotin, khng th th hai l khng th n dng c hiu khng nti-CCP nh du ruthenium (cht c kh nng pht quang) to thnh phc hp min dch kiu sandwich. Cng pht quang t l thun vi nng nti-CCP c trong mu th.
II. CHUN B
1. Ngi thc hin
01 cn b i hc, 01 k thut vin chuyn ngnh ha sinh
2. Phng tin, ha cht
+ Phng tin: My xt nghim nh Cobas e411, e170. e601, rchitect.
+ Ha cht: Ha cht xt nghim nti-CCP, cht chun nti-CCP, cht kim tra cht lng nti-CCP.
3. Ngi bnh
Ngi bnh cn c gii thch v mc ch ca vic ly mu lm xt nghim. Khng s dng cc thuc c Biotin t nht 8 gi trc khi ly mu.
4. Phiu xt nghim
Phiu xt nghim cn ghi y thng tin v tn, tui, gii tnh, khoa phng, chn on ca ngi bnh v ghi r ch nh xt nghim
III. CC BC TIN HNH
1. Ly bnh phm
- Ly 3 ml mu tnh mch vo ng khng c cht chng ng hay ng c cht chng ng l Li-Heparin, EDT . Mu khng v hng cu.
- Sau khi ly mu, em ly tm tch ly huyt thanh hoc huyt tng.
- Bnh phm n nh 3 ngy 28C, 1 thng -20C.
- Bnh phm ch r ng 1 ln v phi bnh phm t nhit phng trc khi phn tch. trnh hin tng bay hi, bnh phm, cht chun, cht kim tra cht lng nn phn tch trong vng 2gi.
2. Tin hnh k thut
- My phn tch cn chun b sn sng thc hin phn tch mu: My c ci t chng trnh xt nghim nti-CCP. My c chun vi xt nghim nti-
-
55
CCP. Kt qu kim tra cht lng vi xt nghim nti-CCP t yu cu khng nm ngoi di cho php v khng vi phm lut kim tra cht lng.
-Ngi thc hin phn tch mu nhp d liu v thng tin ngi bnh v ch nh xt nghim vo my phn tch hoc h thng mng (nu c).
- Np mu bnh phm vo my phn tch
- Ra lnh cho my thc hin phn tch mu bnh phm
- i my phn tch mu theo protocol ca my
- Khi c kt qu cn xem xt nh gi kt qu sau in bo co hoc ghi kt qu vo phiu xt nghim tr cho ngi bnh.
IV. NHN NH KT QU
+ Tr s bnh thng:
-
56
Anti-TG l khng th khng TG. Xt nghim nti-TG thng c ch nh trong mt s bnh v tuyn gip nh: vim tuyn gip Hasimoto, ung th gip, Basedow.
Anti-TG c inh lng theo nguyn l min dch cnh tranh s dng cng ngh ha pht quang hay in ha pht quang.
u tin: Anti-TG trong mu th c vi TG nh du biotin ( pha rn) v khng th ca mu bnh phm gn kt vi khng nguyn.
Sau khi thm khng th khng TG nh du ruthenium (cht c kh nng pht quang) v cc vi ht ph streptavidin, phc hp min dch to thnh tr nn gn kt vi pha rn thng qua s tng tc ca biotin v streptavidin. Nh vy, nng nti-TG trong mu th cng cao th phc hp ny cng thp v do vy tn hiu nh sng pht ra t l nghch vi nng anti-TG c trong mu th.
II. CHUN B
1. Ngi thc hin
01 cn b i hc, 01 k thut vin chuyn ngnh ha sinh.
2. Phng tin, ha cht
+ Phng tin: My xt nghim nh Cobas e411, e170. e601, Architect.
+ Ha cht: Ha cht xt nghim Anti-TG, cht chun Anti-TG, cht kim tra cht lng Anti-TG .
3. Ngi bnh
Ngi bnh cn c gii thch v mc ch ca vic ly mu lm xt nghim. Khng s dng cc thuc c Biotin t nht 8 gi trc khi ly mu.
4. Phiu xt nghim
Phiu xt nghim cn ghi y thng tin v tn, tui, gii tnh, khoa phng, chn on ca ngi bnh v ghi r ch nh xt nghim
III. CC BC TIN HNH
1. Ly bnh phm
- Ly 3 ml mu tnh mch vo ng khng c cht chng ng hay ng c cht chng ng l Na-Heparin v K2,K3-EDTA. Mu khng v hng cu. - - Sau khi ly mu, em ly tm tch ly huyt thanh hoc huyt tng.
- Bnh phm n nh 3 ngy 28C, 1 thng -20C.
- Bnh phm ch r ng 1 ln v phi bnh phm t nhit phng trc khi phn tch. trnh hin tng bay hi, bnh phm, cht chun, cht kim tra cht lng nn phn tch trong vng 2 h.
-
57
2. Tin hnh k thut
- My phn tch cn chun b sn sng thc hin phn tch mu: My c ci t chng trnh xt nghim nti-TG. My c chun vi xt nghim nti-TG. Kt qu kim tra cht lng vi xt nghim nti-TG t yu cu khng nm ngoi di cho php v khng vi phm lut kim tra cht lng.
- Ngi thc hin phn tch mu nhp d liu v thng tin ngi bnh v ch nh xt nghim vo my phn tch hoc h thng mng (nu c).
- Np mu bnh phm vo my phn tch
- Ra lnh cho my thc hin phn tch mu bnh phm
- i my phn tch mu theo protocol ca my
- Khi c kt qu cn xem xt nh gi kt qu sau in bo co hoc ghi kt qu vo phiu xt nghim tr cho ngi bnh.
IV. NHN NH KT QU
- Tr s bnh thng: < 115 IU/mL
- Anti-TG mu tng trong: Mt s bnh v tuyn gip nh: vim tuyn gip Hasimoto, ung th gip, Basedow.
- Anti-TG mu gim trong: S gim nng Anti-TG cng c gi tr theo di hiu qu ca phng php iu tr ca nhng bnh trn.
V. NHNG SAI ST V X TR
- Nhng yu t gy nhiu cho kt qu xt nghim. Kt qu xt nghim khng b nh hng khi:
+ Huyt thanh vng: Bilirubin < 66 mg/dL .
+ Tn huyt: Hemoglobin
-
58
15. NH LNG ANTI-TPO (Anti thyroid peroxydase)
I. NGUYN L
Anti-TPO l khng th khng TPO. Xt nghim nti-TPO thng c ch nh trong Vim tuyn gip Hasimoto. Mt s bnh v tuyn gip nh: vim tuyn gip, Basedow.
Anti-TPO c nh lng theo nguyn l min dch cnh tranh s dng cng ngh ha pht quang hay in ha pht quang.
-
59
u tin: Bnh phm c vi khng th khng TPO nh du phc hp ruthenium.
Sau khi thm TPO nh du biotin v cc vi ht ph streptavidin, khng th khng TPO trong mu cnh tranh vi khng th khng TPO nh du phc hp ruthenium vo v tr gn kt trn khng nguyn TPO nh du biotin. Ton b phc hp tr nn gn kt vi pha rn thng qua s tng tc gia biotin v streptavidin.
Hn hp phn ng c chuyn ti bung o, cc vi ht i t c bt gi ln b mt ca in cc. Nhng thnh phn khng gn kt s b thi ra ngoi bung o bi dung dch ProCell. Cho in p vo in cc s to nn s pht quang ha hc c o bng b khuch i quang t.
II. CHUN B
1. Ngi thc hin
01 cn b i hc, 01 k thut vin chuyn ngnh ha sinh.
2. Phng tin, ha cht
+ Phng tin: My xt nghim nh Cobas e411, e170. e601, Architect.
+ Ha cht: Ha cht xt nghim Anti-TPO, cht chun Anti-TPO, cht kim tra cht lng Anti-TPO .
3. Ngi bnh
Ngi bnh cn c gii thch v mc ch ca vic ly mu lm xt nghim.
4. Phiu xt nghim
Phiu xt nghim cn ghi y thng tin v tn, tui, gii tnh, khoa phng, chn on ca ngi bnh v ghi r ch nh xt nghim
III.CC BC TIN HNH
1. Ly bnh phm
- Ly 3 ml mu tnh mch vo ng khng c cht chng ng hay ng c cht chng ng l NH4,Li, Na-Heparin v K3-EDTA, Natri Citrat, Na fluoride, K Oxalate. Mu khng v hng cu.
- Sau khi ly mu, em ly tm tch ly huyt thanh hoc huyt tng.
- Bnh phm n nh 3 ngy 28C, 1 thng -20C.
- Bnh phm ch r ng 1 ln v phi bnh phm t nhit phng trc khi phn tch. trnh hin tng bay hi, bnh phm, cht chun, cht kim tra cht lng nn phn tch trong vng 2 h.
2.Tin hnh k thut
-
60
- My phn tch cn chun b sn sng thc hin phn tch mu: My c ci t chng trnh xt nghim nti-TPO. My c chun vi xt nghim nti-TPO. Kt qu kim tra cht lng vi xt nghim nti-TPO t yu cu khng nm ngoi di cho php v khng vi phm lut kim tra cht lng.
- Ngi thc hin phn tch mu nhp d liu v thng tin ngi bnh v ch nh xt nghim vo my phn tch hoc h thng mng (nu c).
- Np mu bnh phm vo my phn tch
- Ra lnh cho my thc hin phn tch mu bnh phm
- i my phn tch mu theo protocol ca my
- Khi c kt qu cn xem xt nh gi kt qu sau in bo co hoc ghi kt qu vo phiu xt nghim tr cho ngi bnh.
IV. NHN NH KT QU
- Tr s bnh thng:
-
61
1. Ngi thc hin
01 cn b i hc, 01 k thut vin chuyn ngnh ha sinh. 2. Phng tin, ha cht
2.1.My phn tch sinh ha t ng
MODUL R P800, COB S 6000, U 2700,
2.2.Ha cht - Anti-Apo A1 antibodies
- Tris buffer (pH 7.4)
- Sodioum chlorid
- Polyethylen glycol
- Cht bo qun Ha cht c bo qun 2 80C. Hn s dng: theo ngy ghi trn hp.
3. Ngi bnh: Cn c gii thch v mc ch ca xt nghim 4. Phiu xt nghim: theo quy nh ca b y t, c ch nh xt nghim
ca bc s lm sng. III. CC BC TIN HNH 1. Ly bnh phm
2 ml huyt thanh Huyt thanh n nh trong vng 8 ngy nhit 2-80C, 1 ngy nhit 15-
250C
2. Tin hnh k thut 2.1.Chun b ha cht Chun b ha cht, cht chun, huyt thanh kim tra cht lng xt nghim po A1 2.2.Tin hnh k thut - Ci t chng trnh, cc thng s k thut xt nghim po 1 theo protocol ca my.
- Tin hnh chun po 1. - Kim tra cht lng xt nghim po 1. Nu kt qu kim tra cht lng t (khng vi phm cc lut kim tra cht lng): tin hnh thc hin xt nghim cho ngi bnh; nu kt qu vi phm vo lut kim tra cht lng: chun li my v kim tra cht lng li. - Phn tch mu bnh phm ca ngi bnh theo protocol ca my. - Kt qu sau khi c nh gi s c chuyn vo phn mm qun l d liu hoc vo s lu kt qu (ty thuc vo iu kin ca phng xt nghim). - Tr kt qu cho khoa lm sng, cho ngi bnh
IV. NHN NH KT QU
1. Tr s bnh thng - Nam: 105 175 mg/dl - N: 105 205 mg/dl
-
62
2. Apo A1 gim Apo A1 mu gim l yu t d bo nguy c x va ng mch.
V. NHNG SAI ST V X TR 1. Trc phn tch
Bnh phm v hng cu s nh hng n kt qu xt nghim 2. Trong phn tch
Mu bnh phm ca ngi bnh ch c thc hin khi kt qu kim tra cht lng khng vi phm cc lut ca quy trnh kim tra cht lng; nu khng, phi tin hnh chun v kim tra cht lng li, t mi thc hin xt nghim cho ngi bnh; nu khng t: tin hnh kim tra li cc thng s k thut ca my, sa cha hoc thay mi cc chi tit nu cn. Sau chun v kim tra cht lng li cho t.
3. Sau phn tch Phn tch kt qu thu c vi chn on lm sng, vi kt qu cc xt nghim
khc ca chnh ngi bnh ; nu khng ph hp, tin hnh kim tra li: thng tin trn mu bnh phm, cht lng mu, kt qu kim tra cht lng my, phn tch li mu bnh phm .
-
63
17. NH LNG APO B
I. NGUYN L Min dch o c: Apo B phn ng c hiu vi khng th khng Apo B to
hp cht khng tan lm c mi trng. Mt quang ca mi trng phn ng t l vi nng Apo B trong mu bnh phm.
II. CHUN B
1. Ngi thc hin
01 cn b i hc, 01 k thut vin chuyn ngnh ha sinh. 2. Phng tin, ha cht:
2.1. My phn tch sinh ha t ng
MODUL R P800, COB S 6000, U 2700, 2.2. Ha cht - Anti-Apo B antibodies
- Tris buffer (pH 7.4)
- Sodioum chlorid
- Polyethylen glycol
- Cht bo qun Ha cht c bo qun 2 80C. Hn s dng: theo ngy ghi trn hp.
3. Ngi bnh Cn c gii thch v mc ch ca xt nghim
4. Phiu xt nghim Ghi y cc thng tin tn, tui, gii tnh, khoa, phng, chn on bnh, s
ging..Phiu xt nghim phi c ch nh xt nghim. III. CC BC TIN HNH 1. Ly bnh phm
2 ml huyt thanh Huyt thanh; n nh trong vng 8 ngy nhit 2 - 80C, 1 ngy nhit 15-
250C.
2. Tin hnh k thut 2.1.Chun b ha cht Chun b ha cht, cht chun, huyt thanh kim tra cht lng xt nghim po B 2.2.Tin hnh k thut
- Ci t chng trnh, cc thng s k thut xt nghim po B theo protocol ca my.
- Tin hnh chun po B. - Kim tra cht lng xt nghim po B. Nu kt qu kim tra cht lng t (khng vi phm cc lut kim tra cht lng): tin hnh thc hin xt nghim cho
-
64
ngi bnh; nu kt qu vi phm vo lut kim tra cht lng: chun li my v kim tra cht lng li.
- Phn tch mu bnh phm ca ngi bnh theo protocol ca my. - Kt qu sau khi c nh gi s c chuyn vo phn mm qun l d liu hoc vo s lu kt qu (ty thuc vo iu kin ca phng xt nghim).
- Tr kt qu cho khoa lm sng, cho ngi bnh IV. NHN NH KT QU
1. Tr s bnh thng - Nam: 60 - 140 mg/dl
- N: 66 - 130 mg/dl
2. Apo B tng po B mu tng l yu t d bo nguy c x va ng mch.
V. NHNG SAI ST V X TR 1. Trc phn tch
Bnh phm v hng cu s nh hng n kt qu xt nghim 2. Trong phn tch
Mu bnh phm ca ngi bnh ch c thc hin khi kt qu kim tra cht lng khng vi phm cc lut ca quy trnh kim tra cht lng; nu khng, phi tin hnh chun v kim tra cht lng li, t mi thc hin xt nghim cho ngi bnh; nu khng t: tin hnh kim tra li cc thng s k thut ca my, sa cha hoc thay mi cc chi tit nu cn. Sau chun v kim tra cht lng li cho t.
3. Sau phn tch Phn tch kt qu thu c vi chn on lm sng, vi kt qu cc xt nghim
khc ca chnh ngi bnh ; nu khng ph hp, tin hnh kim tra li: thng tin trn mu bnh phm, cht lng mu, kt qu kim tra cht lng my, phn tch li mu bnh phm .
-
65
18. NH LNG AFP ( Alpha fetoprotein)
FP l glycoprotein trng lng phn t 70 000 Daltons, c sn xut bi ti non, t bo gan cha bit ha v ng tiu ha ca bo thai. Xt nghim FP thng c ch nh trong bnh ung th gan nguyn pht, x gan v theo di trong iu tr.
I. NGUYN L
AFP c nh lng bng phng php min dch sandwich s dng cng ngh ha pht quang hay in ha pht quang. AFP c trong mu th ng vai tr khng nguyn c kp gia hai khng th, khng th th nht l khng th n dng c hiu khng AFP nh du biotin, khng th th hai l khng th n dng c hiu khng AFP nh du ruthenium (cht c kh nng pht quang) to thnh phc hp min dch kiu sandwich. Cng pht quang t l thun vi nng AFP c trong mu th.
II. CHUN B
1. Ngi thc hin: 01 cn b i hc, 01 k thut vin chuyn ngnh hoa sinh
2. Phng tin, ha cht
- Phng tin: My xt nghim nh Cobas e411, e170. e601, Architect.
- Ha cht: Ha cht xt nghim AFP, cht chun AFP, cht kim tra cht lng AFP.
3. Ngi bnh: Ngi bnh cn c gii thch v mc ch ca vic ly mu lm xt nghim.
4. Phiu xt nghim: Phiu xt nghim cn ghi y thng tin v tn, tui, gii tnh, khoa phng, chn on ca ngi bnh v ghi r ch nh xt nghim
III.CC BC TIN HNH
1. Ly bnh phm
- Ly 3 ml mu tnh mch vo ng khng c cht chng ng hay ng c cht chng ng l Li, Na-Heparin v K3-EDTA v Natri Citrat. Mu khng v hng cu.
- Sau khi ly mu, em ly tm tch ly huyt thanh hoc huyt tng.
- Bnh phm n nh 7 ngy 28C, 3 thng -20C. Bnh phm ch r ng 1 ln v phi bnh phm t nhit phng trc khi phn tch. trnh hin tng bay hi, bnh phm, cht chun, cht kim tra cht lng nn phn tch trong vng 2gi.
2. Tin hnh k thut
-
66
- My phn tch cn chun b sn sng thc hin phn tch mu: My c ci t chng trnh xt nghim FP. My c chun vi xt nghim FP. Kt qu kim tra cht lng vi xt nghim FP t yu cu khng nm ngoi di cho php v khng vi phm lut kim tra cht lng.
- Ngi thc hin phn tch mu nhp d liu v thng tin ngi bnh v ch nh xt nghim vo my phn tch hoc h thng mng (nu c).
- Np mu bnh phm vo my phn tch
- Ra lnh cho my thc hin phn tch mu bnh phm
- i my phn tch mu theo protocol ca my
- Khi c kt qu cn xem xt nh gi kt qu sau in bo co hoc ghi kt qu vo phiu xt nghim tr cho ngi bnh.
IV. NHN NH KT QU
- Tr s bnh thng: < 7.0 ng/ml
-FP mu tng trong: Ung th gan nguyn pht c mc tng cao nht, Ung th gan th pht mc tng t hn c v tn sut v nng , U nguyn bo phi, Mt s
bnh gan nh vim gan, x gan.,FP cng vi HCG v uE3 l b ba xt nghim dng cho chn on trc sinh i vi bnh Down v d tt bnh sinh nh tt nt t sng
V. NHNG SAI ST V X TR
- Nhng yu t gy nhiu cho kt qu xt nghim. Kt qu xt nghim khng b nh hng khi:
+ Huyt thanh vng: Bilirubin < 65 mg/dL hay 1112 mol/L.
+ Tn huyt: Hemoglobin
-
67
19. O HOT ALT
(Alanin transaminase)
LT cn c gi l GPT (Glutamat pyruvat transaminase)
o hot LT thng c lm cng vi ST xc nh bnh l v gan, theo di tin trin ca bnh. Ngoi ra LT cng c phi hp vi mt s xt nghim khc nh GGT theo di ngi bnh nghin ru.
I. NGUYN L
Hot ca enzym ALT trong mu ca ngi bnh c xc nh theo phng php ng hc enzyme da trn phn ng:
L.Alanin + -cetoglutarat ALT L.Glutamat + Pyruvat
LDH
Pyruvat + NADH +H+
L- lactate+ NAD+
Hot LP c o bng s gim nng N DH bc song 340 nm theo thi gian.
II. CHUN B
1. Ngi thc hin
02 cn b l bc s v k thut vin c o to v chuyn ngnh ha sinh.
2. Phng tin, ha cht
2.1. Phng tin
- H thng my phn tch ha sinh ca hng Roche (MODUL R, COB S 6000, COBAS 8000), hng Olympus (AU 640, AU 2700, AU5800).
- My ly tm
- Cc ng xt nghim c chng ng bng Li-Heparin hoc EDT hoc khng chng ng.
- Pipt t ng cc loi 1000l, 500 l, 100l, 50 l v 10 l.
- u cn tng ng cc loi pipet t ng.
- Bng, cn, kim ly mu, gi ng bnh phm.
-
68
- Bn ly mu.
- Gng tay
2.2. Ho cht
- Ho cht lm xt nghim ALP ca hng ROCHE, OLYMPUS.
- Huyt thanh kim tra ca BIO-RAD.
2.3. Bnh phm
- Ly 3 ml mu tnh mch cho vo ng chng ng bng Li-Heparin, EDT , hoc ng khng chng ng
- Ly tm tch huyt tng hoc huyt thanh
- Mu bnh phm cn c phn tch cng sm cng tt. C th bo qun mu huyt thanh hoc huyt tng 7 ngy nhit 2-80C.
3. Ngi bnh: c t vn xt nghim, chun b t tng khi khm bnh, nhn n sng ly mu.
4. Phiu xt nghim: in y thng tin v ngi bnh theo quy nh. Phiu xt nghim c ch nh xt nghim ALT trong mu.
III. CC BC TIN HNH
- Ci chng trnh trn my theo protocol ca my: ch lm khi bt u trin khai xt nghim trn my v khi c thay i trong chng trnh ci t.
- Dng ng chun: c lm khi bt u trin khai xt nghim trn my, khi thay i mt trong cc yu t: nng chun mi, thuc th mi, thay bng n hay thay cng phn ng, v khi thy kt qu kim tra cht lng khng t.
- Mu huyt thanh kim tra cht lng, mu bnh phm o hot LP c phn tch trn my phn tch sinh ho t ng MODUL R, COB S 6000, COB S 8000 (hng Roche), hoc cc my U 640, U 2700, U 5800 (hng Minh Tm) theo protocol ca my.
- Mu bnh phm ch c chy trn my phn tch khi kt qu kim tra cht lng t c chnh xc v xc thc trong gii hn cho php v khng vi phm cc lut ca quy trnh kim tra cht lng.
- Kt qu sau khi c nh gi s c in vo phiu xt nghim, in vo s lu tr hoc c chuyn vo phn mm qun l d liu in ra bng my.
IV. NHN NH KT QU
1. Tr s bnh thng
Nam: < 41 U/L.
-
69
N:
-
70
ST cn c gi l GOT (Glutamat oxaloacetat transaminase)
I. NGUYN L
o hot ST thng c lm cng vi LT xc nh bnh l v theo di tin trin ca gan hay tim mch,. Ngoi ra ST cng c phi hp vi mt s xt nghim khc nh GGT theo di ngi bnh nghin ru.
Hot ca enzym AST trong mu ca ngi bnh c xc nh theo phng php ng hc enzyme, theo phn ng:
L-aspartat + -cetoglutarat AST Glutamat + Oxaloacetat
GOT
MDH
Oxaloacetat + NADH + H+
L-malate + NAD+
Hot ST c o bng s gim nng N DH theo thi gian bc sng 340 nm.
II. CHUN B
1. Ngi thc hin
01bc s v 01 k thut vin c o to v chuyn ngnh ha sinh.
2. Phng tin, ha cht
2.1. Phng tin
- H thng my phn tch ha sinh ca hng Roche (MODUL R, COB S 6000, COBAS 8000), hng Olympus (AU 640, AU 2700, AU5800).
- My ly tm
- Cc ng xt nghim c chng ng bng Li-Heparin hoc EDT hoc khng chng ng.
- Pipt t ng cc loi 1000l, 500 l, 100l, 50 l v 10 l.
- u cn tng ng cc loi pipet t ng.
- Bng, cn, kim ly mu, gi ng bnh phm.
- Bn ly mu.
- Gng tay
2.2. Ho cht
+ Ho cht lm xt nghim AST ca hng ROCHE, OLYMPUS.
+ Huyt thanh kim tra ca BIO-RAD.
-
71
+ Chun ca ST.
2.3. Bnh phm
- Ly 3 ml mu tnh mch vo ng chng ng bng Li-Heparin, EDT , hoc ng khng chng ng
- Ly tm tch huyt tng hoc huyt thanh
- Mu bnh phm cn c phn tch cng sm cng tt. C th bo qun mu huyt thanh hoc huyt tng 7 ngy nhit 2-80C.
3. Ngi bnh: c t vn xt xt nghim, chun b t tng khi khm bnh, nhn n sng ly mu.
4. Phiu xt nghim: in y thng tin v ngi bnh theo quy nh. Phiu xt nghim c ch nh xt nghim AST trong mu.
III. CC BC TIN HNH
- Ci chng trnh trn my theo protocol ca my: ch lm khi bt u trin khai xt nghim trn my v khi c thay i trong chng trnh ci t.
- Dng ng chun: c lm khi bt u trin khai xt nghim trn my, khi thay i mt trong cc yu t: nng chun mi, thuc th mi, thay bng n hay thay cng phn ng, v khi thy kt qu kim tra cht lng khng t.
- Mu huyt thanh kim tra cht lng, mu bnh phm o hot ST c phn tch trn my phn tch sinh ho t ng MODUL R, COB S 6000, COB S 8000 (hng Roche), hoc cc my U 640, U 2700, U 5800 (hng Minh Tm) theo protocol ca my.
- Mu bnh phm ch c chy trn my phn tch khi kt qu kim tra cht lng t c chnh xc v xc thc trong gii hn cho php v khng vi phm cc lut ca quy trnh kim tra cht lng.
- Kt qu sau khi c nh gi s c in vo phiu xt nghim, in vo s lu tr hoc c chuyn vo phn mm qun l d liu in ra bng my.
VI. NHN NH KT QU
Tr s bnh thng:
Nam: < 37 U/L.
N: < 31 U/L .
- AST mu tng trong cc nguyn nhn:
Cc bnh gan (t s ST/ALT
-
72
Cc bnh gan (t s ST/ LT >1): X gan, Vim gan do ru, Xm nhim gan ( do di cn ung th, nhim sarcoid, lao, u lympho, luput ban ).
Cc bnh v tim: suy tim mt b ( gan xung huyt), vim c tim, nhi mu c tim , bp tim ngoi lng ngc, phu thut tim, sau thng tim (t s AST/ALT>1).
Vim ti mt.
Nhim c ru cp.
Vim tu cp hoi t.
Vim a c, vim da v c,
Hi chng vi lp.
- Hot AST c th gim trong cc nguyn nhn chnh sau:
Bnh Beriberi.
Nhim toan ceton do i tho ng.
Lc mu.
C thai
Hi chng ure mu cao.
VII. NHNG SAI ST V X TR
- Khi thy kt qu AST bt thng (cao hn hoc thp hn gi tr bnh thng) cn kim tra li kt qu bng cch:
+ Nhc ng mu kim tra xem c ng dy hoc bt thng v mu sc huyt tng hay khng?
+ Kim tra li thng tin ng mu, i chiu vi thng tin trn phiu yu cu xt nghim: h tn ngi bnh, tui, ging, khoa
Nu thy khng c g bt thng, nn chy li kim tra li ln na trn my cng phi hp vi mu huyt thanh kim tra hoc chuyn sang my khc.
- Cc yu t gp phn lm thay i kt qu xt nghim:
+ Mu mu b v hng cu
+ Cc thuc c th lm tng hot ST l: cetaminophen, allopurinol, mt s loi khng sinh, acid ascorbic, chlpropamid, cholestyramin, cholinergic, clofibrat, codein, statin, hydralazin, isoniazid, meperidin, methyldopa, morphin,
thuc nga thai ung, phenothiazin, procainamid, pyridoxin, salicylat, sufonamid, verapamil, vitamin A.
+ Cc thuc c th lm gim hot ST l; metronidazol, trifluoperazin.
-
73
21. NH LNG 1 ACID GLYCOPROTEIN
I. NGUYN L
1 cid glycoprotein l mt protein pha cp. Xt nghim 1 Acid glycoprotein thng c s dng trong cc trng hp vim, nhim trng.
-
74
1 Acid glycoprotein trong mu ca ngi bnh c xc nh theo phng php min dch o c.
Khng th khng 1 cid glycoprotein trong thuc th kt hp vi