evaluasi & tindak lanjut pme immunolgi - dr farida oesman, sppk(k)

8/16/2019 Evaluasi & Tindak Lanjut PME Immunolgi - Dr Farida Oesman, SpPK(K)

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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


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Farida OesmanDepartment of Clinical Pathology

Faculty of Medicine, University of Indonesia


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POST ANALYTIC

ANALYTIC

PRA ANALYTIC

QUALITY

CONTROL

QUALITY

ASSURANCE


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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


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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


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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


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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


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PRE ANALYTIC

RIGHT

• INVESTIGATION

• SAMPLE

• COLLECTION

• TECHNIQUE

• LABELING

• QUANTITY

• TRANSPORTATION


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POST ANALYTIC

• ACCURATE RECORDING

• RANGE OF NORMAL VALUE

• SEX AGE RELATED

VARIATION

• TURNAROUND TIME


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IMMUNOLOGY

ASSAYS

ARE PERFORMED TO DETECT

• ANTIGEN

• ANTIBODY AGAINST ANTIGEN

IN BODY FLUID

BASED ON

ANTIGEN AND ANTIBODY REACTION


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ANTIGEN ANTIBODY

REACTION

PRIMARY SECONDARY

+

ANTIGEN ANTIBODY Ag-Ab COMPLEX

USING LABEL

MATERIAL


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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


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ANTIBODY

ANTIGEN


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ENZYME

RADIO

ACTIVE

FLUORESCENT

EIA

RIA

IFA

• COMPETITIVE

• NON COMPETITIVE

INDIRECT

SANDWICH

• DIRECT

• INDIRECT


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REQUIRE

SEPARATION OF

BOUND Ag – Ab

WASHING

MEASUREMENT

OF SMALL

ANALYTES

IMMUNOASSAYS

METHOD

HOMOGENEOUS

HETEROGENEOUS

NOT REQUIRE

SEPARATION OF

BOUND Ag – Ab

EASY & FAST


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HOMOGENEOUSMETHOD

HOTEROGENEOUSMETHOD


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IMMUNOASSAYS

METHOD

COMPETITIVE

NON COMPETITIVE

HIGH

SENSITIVE

SPECIFIC

MEASUREMENT

OF ANTIGEN

MEASUREMENT

OF CRITICAL

ANALYTES


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ONE STEP

LIMITED AMOUNT OF ANTIBODY

AMOUNT OF ANALYTES

IS INVERSELY PROPORTIONAL

TO THE AMOUNT OF SIGNAL

COMPETITIVE


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COMPETITIVE

TWO STEP

EXCESS OF ANTIBODY

IMPROVED ASSAY SENSITIVITY


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AMOUNT OF ANALYTES IS DIRECTLY PROPORTIONAL

TO THE AMOUNT OF SIGNAL

ANTIBODIES BIND TO TWO SITES ON ANALYTE

SANDWICH


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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


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IMMUNOASSAYS

ASSAY INTERFERENCE

• ANALYTES EXCESS

• HUMAN ANTI-MOUSE ANTIBODIES

• HETEROPHILE ANTIBODIES

ONE STEP IMMUNOASSAY CAN BE

PRONE TO INTERFERENCES THAT

EFFECT SENSITIVITY SPECIFICITY


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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


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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 (+)


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Weak sample: conjugate reacts withall available binding sites on thetarget antibody.

Prozone with simultaneous

sandwich assay


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Strong sample: antibody binds all theconjugate and all the solid phasesites.

Prozone with simultaneoussandwich assay


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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 (-)


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IN IMMUNOASSAY THAT UTILIZE

MOUSE MONOCLONAL ANTIBODIES


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IN IMMUNOASSAY THAT UTILIZE

MOUSE MONOCLONAL ANTIBODIES


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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


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SECONDARY

Ag Ab REACTION

• AGGLUTINATION

DIRECT

INDIRECT

INHIBITION

• PRECIPITATION

• FLOCULATION

• COMPLEMENT FIXATION


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LATTICE FORMATION

AGGLUTINATION

Ag Ab

EQUIVALENT


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PROZONE

NO AGGLUTINATION

FALSE NEGATIVE

DILUTE SAMPLE

Ab EXCESS


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Ag EXCESS

DISSOCIATION OF Ag-Ab COMPLEX

NO AGGLUTINATION

FALSE NEGATIVE

DILUTE SAMPLE

POSTZONE


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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


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evaluasi & tindak lanjut pme immunolgi - dr farida oesman, sppk(k)

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