Immunoassay Precipitation Manual Double immunodiffusion Automation Turbidimetry nephelometry

면역측정법 (Immunoassay)의 종류 • Precipitation(침강법) – Manual: Double immunodiffusion – Automation: Turbidimetry, nephelometry • Agglutination(응집법) – Manual: Latex agglutination, hemagglutination – Automation: Turbidimetry, nephelometry • EIA (효소면역측정법) – Colorimetric EIA – Fluorescent EIA – Chemiluminescent EIA • FIA (fluorescent immunoassay, 형광면역측정법) • CLIA (chemiluminescent immunoassay, 화학발광면역측 정법)

효소면역측정법(Enzyme immunoassay, EIA) – 효소를 표지자로 이용 – 효소 + 기질 반응 – 발색량을 분광광도계로 측정 – Alkaline phosphatase, horseradish peroxidase, B -galactosidase – Homegeneous EIA : enzyme multiplied immunoassay technique(EMIT) – Heterogeneous EIA : • Competitive • Non-competitive – 바이러스 항원, 항체, 종양 표지자 및 각종호르몬 의 측정

Competitive Enzyme immuno assay

Non-competitive EIA

Fluorescence immunoassay • 원리 : fluorescent isothionate(FITC), 형광물질 표지

Mononuclear cell isolation • Ficoll-Hypaque (FH) separation – 두가지 물질을 섞어 비중을 1. 077에 맞춤 – Centrifuge후 lymphocyte는 plasma-Ficoll. Hypaque interface에 위치 • Cell viability – > 80%, minimal – > 90%, preferable

Human Leukocyte Antigen (HLA) • Human Major Histocompatibility Complex (MHC) HLA 6 p 21. 3 Typing Methods Serological Analysis Microlymphocytotoxicity Molecular Analysis PCR-SSP Reverse SSOP PCR-SBT

1 (negative, 1 -10%) 8 8 (strong positive, 81 -100%)

PCR-SSOP Reverse SSOP B*44/B*55 PCR-SSP

알레르기 질환의 검사 • 1) 혈청 총 Ig. E 측정 • 2) allergen specific Ig. E antibody – – 피부반응검사 RAST MAST CAP system

Typical Serological Profile After Acute Infection Note that during reinfection, Ig. M may be absent or present at a low level transientl

바이러스성 간염 • Hepatitis A – Anti-HAV Ig. M, HAV항원 : 대변, PCR • • • HEV : Anti-HEV, PCR HBV HCV HDV Epstein-Barr바이러스에 의한 전염성 단핵 구증

ELISA for HIV antibody Microplate ELISA for HIV antibody: colored wells indicate reactivity

Western Blot