Evaluasi & Tindak Lanjut PME Immunolgi - Dr Farida Oesman, SpPK(K)
-
Upload
steviodorgipratama -
Category
Documents
-
view
278 -
download
0
Transcript of Evaluasi & Tindak Lanjut PME Immunolgi - Dr Farida Oesman, SpPK(K)
-
8/16/2019 Evaluasi & Tindak Lanjut PME Immunolgi - Dr Farida Oesman, SpPK(K)
1/40
Nama : dr. Farida Oesman, SpPK(K)
Tempat, tgl lahir : Yogyakarta, 4 April 1951
Departemen : Patologi Klinik FKUI – RSCM
Riwayat pendidikan :
S1: FKUI, 1975
Sp1: Spesialis Patologi Klinik FKUI, 1983
Sp2: Konsultan Alergi Imunologi, 2007
Jabatan saat ini:
Ketua Divisi Alergi Imunologi Departemen Patologi KlinikFKUI-RSCM
Kepala Laboratorium Imunologi Dept Patologi Klinik RSCM
Staf Pengajar Departemen Patologi Klinik FKUI
CURRICULUM VIT E
-
8/16/2019 Evaluasi & Tindak Lanjut PME Immunolgi - Dr Farida Oesman, SpPK(K)
2/40
Farida OesmanDepartment of Clinical Pathology
Faculty of Medicine, University of Indonesia
-
8/16/2019 Evaluasi & Tindak Lanjut PME Immunolgi - Dr Farida Oesman, SpPK(K)
3/40
POST ANALYTIC
ANALYTIC
PRA ANALYTIC
QUALITY
CONTROL
QUALITY
ASSURANCE
-
8/16/2019 Evaluasi & Tindak Lanjut PME Immunolgi - Dr Farida Oesman, SpPK(K)
4/40
-
8/16/2019 Evaluasi & Tindak Lanjut PME Immunolgi - Dr Farida Oesman, SpPK(K)
5/40
QUALITY
ASSURANCE
THE RIGHT TEST IS CARRIED ON
THE RIGHT SAMPLE FROM
THE RIGHT PERSON WITH
THE RIGHT RESULT INTERPRETATION
IS DELIVERED TO
THE RIGHT PERSON THE RIGHT TIME
OVERALL PROCESS THAT
ENSURE THAT FINAL RESULTS
REPRTED BY LABORATORY ARE CORRECT
-
8/16/2019 Evaluasi & Tindak Lanjut PME Immunolgi - Dr Farida Oesman, SpPK(K)
6/40
QUALITY
CONTROL
IS A STATISTICAL PROCESS USED TO
MONITOR & EVALUATE THE ANALYTICAL PROCESS
THAT PRODUCES PATIENT RESULTS
TO ENSURE THAT THE RESULTS
GENERATED BY TEST ARE CORRECT
MUST BE INCLUDED DURING EACH ASSAY RUN
TO VERIVY THE TEST IS WORKING PROPERLY
-
8/16/2019 Evaluasi & Tindak Lanjut PME Immunolgi - Dr Farida Oesman, SpPK(K)
7/40
-
8/16/2019 Evaluasi & Tindak Lanjut PME Immunolgi - Dr Farida Oesman, SpPK(K)
8/40
QUALITY
ASSURANCE
GOOD LABORATORY PRACTICES
• PROPER COLLECTION OF SAMPLES
• APPROPRIATE IDENTIFICATION OF SPECIMEN
• PROMPT TRANSPORTATION TO LABORATORY
AT APPROPIATE TEMPERATURE
• SAMPLE PREPARATION STORAGE UNDER
CONDITION WHICH PREVENT DETERIORATION
• ACCURATE PERFORMANCE TEST
• RELEASE REPORT AFTER PROPER SCRUTINY
• DELIVERY OF REPORTS TO CORRECT
DESTINATION IN SHORTEST POSSIBLE TIME
-
8/16/2019 Evaluasi & Tindak Lanjut PME Immunolgi - Dr Farida Oesman, SpPK(K)
9/40
ANALYTIC
• EQUIPMENT RELIABILITY
• REAGENT STABILITY, INTEGRITY
AND EFFICIENCY
• ADEQUATE CALIBRATION
• RIGHT TECHNIQUE
• PROCEDURAL REALIBILITY USING SOP
• PROFIENCY OF PERSONNEL
• INTERNAL QC
• EXTERNAL QA
ACCURACY PRECISSION
-
8/16/2019 Evaluasi & Tindak Lanjut PME Immunolgi - Dr Farida Oesman, SpPK(K)
10/40
PRE ANALYTIC
RIGHT
• INVESTIGATION
• SAMPLE
• COLLECTION
• TECHNIQUE
• LABELING
• QUANTITY
• TRANSPORTATION
-
8/16/2019 Evaluasi & Tindak Lanjut PME Immunolgi - Dr Farida Oesman, SpPK(K)
11/40
POST ANALYTIC
• ACCURATE RECORDING
• RANGE OF NORMAL VALUE
• SEX AGE RELATED
VARIATION
• TURNAROUND TIME
-
8/16/2019 Evaluasi & Tindak Lanjut PME Immunolgi - Dr Farida Oesman, SpPK(K)
12/40
IMMUNOLOGY
ASSAYS
ARE PERFORMED TO DETECT
• ANTIGEN
• ANTIBODY AGAINST ANTIGEN
IN BODY FLUID
BASED ON
ANTIGEN AND ANTIBODY REACTION
-
8/16/2019 Evaluasi & Tindak Lanjut PME Immunolgi - Dr Farida Oesman, SpPK(K)
13/40
ANTIGEN ANTIBODY
REACTION
PRIMARY SECONDARY
+
ANTIGEN ANTIBODY Ag-Ab COMPLEX
USING LABEL
MATERIAL
-
8/16/2019 Evaluasi & Tindak Lanjut PME Immunolgi - Dr Farida Oesman, SpPK(K)
14/40
ANTIGEN ANTIBODY
REACTION
DEPENDS ON ANTIBODY
• AFFINITY
THE STRENGTH OF MONOVALENT
ANTIGEN AND ANTIBODY BOND
•
AVIDITY
THE STRENGTH OF MULTIVALENT
ANTIGEN AND ANTIBODY BOND
• SPECIFICITY
-
8/16/2019 Evaluasi & Tindak Lanjut PME Immunolgi - Dr Farida Oesman, SpPK(K)
15/40
-
8/16/2019 Evaluasi & Tindak Lanjut PME Immunolgi - Dr Farida Oesman, SpPK(K)
16/40
ANTIBODY
ANTIGEN
-
8/16/2019 Evaluasi & Tindak Lanjut PME Immunolgi - Dr Farida Oesman, SpPK(K)
17/40
ENZYME
RADIO
ACTIVE
FLUORESCENT
EIA
RIA
IFA
• COMPETITIVE
• NON COMPETITIVE
INDIRECT
SANDWICH
• DIRECT
• INDIRECT
-
8/16/2019 Evaluasi & Tindak Lanjut PME Immunolgi - Dr Farida Oesman, SpPK(K)
18/40
REQUIRE
SEPARATION OF
BOUND Ag – Ab
WASHING
MEASUREMENT
OF SMALL
ANALYTES
IMMUNOASSAYS
METHOD
HOMOGENEOUS
HETEROGENEOUS
NOT REQUIRE
SEPARATION OF
BOUND Ag – Ab
EASY & FAST
-
8/16/2019 Evaluasi & Tindak Lanjut PME Immunolgi - Dr Farida Oesman, SpPK(K)
19/40
HOMOGENEOUSMETHOD
HOTEROGENEOUSMETHOD
-
8/16/2019 Evaluasi & Tindak Lanjut PME Immunolgi - Dr Farida Oesman, SpPK(K)
20/40
IMMUNOASSAYS
METHOD
COMPETITIVE
NON COMPETITIVE
HIGH
SENSITIVE
SPECIFIC
MEASUREMENT
OF ANTIGEN
MEASUREMENT
OF CRITICAL
ANALYTES
-
8/16/2019 Evaluasi & Tindak Lanjut PME Immunolgi - Dr Farida Oesman, SpPK(K)
21/40
ONE STEP
LIMITED AMOUNT OF ANTIBODY
AMOUNT OF ANALYTES
IS INVERSELY PROPORTIONAL
TO THE AMOUNT OF SIGNAL
COMPETITIVE
-
8/16/2019 Evaluasi & Tindak Lanjut PME Immunolgi - Dr Farida Oesman, SpPK(K)
22/40
COMPETITIVE
TWO STEP
EXCESS OF ANTIBODY
IMPROVED ASSAY SENSITIVITY
-
8/16/2019 Evaluasi & Tindak Lanjut PME Immunolgi - Dr Farida Oesman, SpPK(K)
23/40
AMOUNT OF ANALYTES IS DIRECTLY PROPORTIONAL
TO THE AMOUNT OF SIGNAL
ANTIBODIES BIND TO TWO SITES ON ANALYTE
SANDWICH
-
8/16/2019 Evaluasi & Tindak Lanjut PME Immunolgi - Dr Farida Oesman, SpPK(K)
24/40
SOLID PHASE
SIZE SHAPE ARE CRITICAL FACTORS
CAPTURING CAPACITY OF
ANTIGEN OR ANTIBODY
• INNER WALL OF TUBE, MICROTITER WELL
RELATIVELY LARGE
• MAGNETIC OR LATEX PARTICLES
LARGER TOTAL SURFACE AREA
SHORTEN IMMUNOLOGIC REACTION
IMMOBILIZATION OF ANTIGENS / ANTIBODIES BY
COVALENT BINDING OR PHYSICAL ADSORPTION
-
8/16/2019 Evaluasi & Tindak Lanjut PME Immunolgi - Dr Farida Oesman, SpPK(K)
25/40
IMMUNOASSAYS
ASSAY INTERFERENCE
• ANALYTES EXCESS
• HUMAN ANTI-MOUSE ANTIBODIES
• HETEROPHILE ANTIBODIES
ONE STEP IMMUNOASSAY CAN BE
PRONE TO INTERFERENCES THAT
EFFECT SENSITIVITY SPECIFICITY
-
8/16/2019 Evaluasi & Tindak Lanjut PME Immunolgi - Dr Farida Oesman, SpPK(K)
26/40
FALSELY DECREASED LEVEL OF ANALYTES
PROZONE
(HOOK EFFECT)
• HIGH LEVEL OF ANALYTES
• LIMITED QUANTITIES OF
CAPTURE OR DETECTION
ANTIGEN / ANTIBODY
LIKELY TO BE OBSERVED IN
ONE STEP SANDWICH METHOD
-
8/16/2019 Evaluasi & Tindak Lanjut PME Immunolgi - Dr Farida Oesman, SpPK(K)
27/40
-
8/16/2019 Evaluasi & Tindak Lanjut PME Immunolgi - Dr Farida Oesman, SpPK(K)
28/40
1. Antigen and conjugate
together in the well
3. Wash and then add substate.
Colour
2. Add the sample. If specific antibodies are present
they will bind to both plate and conjugate.
PROZONE (+)
-
8/16/2019 Evaluasi & Tindak Lanjut PME Immunolgi - Dr Farida Oesman, SpPK(K)
29/40
Weak sample: conjugate reacts withall available binding sites on thetarget antibody.
Prozone with simultaneous
sandwich assay
-
8/16/2019 Evaluasi & Tindak Lanjut PME Immunolgi - Dr Farida Oesman, SpPK(K)
30/40
Strong sample: antibody binds all theconjugate and all the solid phasesites.
Prozone with simultaneoussandwich assay
-
8/16/2019 Evaluasi & Tindak Lanjut PME Immunolgi - Dr Farida Oesman, SpPK(K)
31/40
1. Antigen on the micro plate
2. Add the sample. If specific antibodies are present
they will bind.
3. Wash and then addconjugated antigen.
4. Wash and then add substate.
Colour
PROZONE (-)
-
8/16/2019 Evaluasi & Tindak Lanjut PME Immunolgi - Dr Farida Oesman, SpPK(K)
32/40
IN IMMUNOASSAY THAT UTILIZE
MOUSE MONOCLONAL ANTIBODIES
-
8/16/2019 Evaluasi & Tindak Lanjut PME Immunolgi - Dr Farida Oesman, SpPK(K)
33/40
IN IMMUNOASSAY THAT UTILIZE
MOUSE MONOCLONAL ANTIBODIES
-
8/16/2019 Evaluasi & Tindak Lanjut PME Immunolgi - Dr Farida Oesman, SpPK(K)
34/40
MINIMIZED BY
HAMA
HETEROPHILE Ab
•USING TWO STEP IMMUNOASSAY
EXTRA WASH STEP WILL WASH AWAY
THE INTERFERENCE
• ADDING BLOCKING SUBSTANCES
BIND TO HAMA / HETEROPHILE Ab
• DILUTION OF P TIENT’S SPECIMEN
DECREASE LEVEL OF HAMA /
HETEROPHILE Ab
-
8/16/2019 Evaluasi & Tindak Lanjut PME Immunolgi - Dr Farida Oesman, SpPK(K)
35/40
SECONDARY
Ag Ab REACTION
• AGGLUTINATION
DIRECT
INDIRECT
INHIBITION
• PRECIPITATION
• FLOCULATION
• COMPLEMENT FIXATION
-
8/16/2019 Evaluasi & Tindak Lanjut PME Immunolgi - Dr Farida Oesman, SpPK(K)
36/40
LATTICE FORMATION
AGGLUTINATION
Ag Ab
EQUIVALENT
-
8/16/2019 Evaluasi & Tindak Lanjut PME Immunolgi - Dr Farida Oesman, SpPK(K)
37/40
PROZONE
NO AGGLUTINATION
FALSE NEGATIVE
DILUTE SAMPLE
Ab EXCESS
-
8/16/2019 Evaluasi & Tindak Lanjut PME Immunolgi - Dr Farida Oesman, SpPK(K)
38/40
Ag EXCESS
DISSOCIATION OF Ag-Ab COMPLEX
NO AGGLUTINATION
FALSE NEGATIVE
DILUTE SAMPLE
POSTZONE
-
8/16/2019 Evaluasi & Tindak Lanjut PME Immunolgi - Dr Farida Oesman, SpPK(K)
39/40
IMMUNOASSAY
ACCURACY PRECISSION
• METHOD
COMPETITIVE / NON COMPETITIVE
ONE STEP / TWO STEP
• INTERFERENCE
HIGH DOSE HOOK EFFECT (PROZONE)
HAMA / HETEROPHILE Ab
• THE NUMBER OF Ag Ab
-
8/16/2019 Evaluasi & Tindak Lanjut PME Immunolgi - Dr Farida Oesman, SpPK(K)
40/40