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

ELISA

Competitive Enzyme immuno assay Non-competitive EIA

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

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

HLA class I and II molecules Class I • Present on all nucleated cells • HLA-A, HLA-B, or HLA-C Class II • B lymphocytes, activated T lymphocytes, monocytes, macrophages, Langerhans cells, dendritic cells, endothelium, and epithelial cells Structures of HLA class I and II molecules J Allergy Clin Immunol. 2010 Feb; 125(2 Suppl 2): S 324 -35

Microlymphocytotoxicity test • HLA 항혈청 +림프구 부유액 +보체 • HLA 항원 존재시 항원-항체반응 → 세포 융해 • Eosin Y → Inverted contrast microscope

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

HLA crossmatch • Detection of Ab in recipient blood for HLA Ag from the donor • Complement-dependent cytotoxicity (CDC) – Sensitivity : Basic NIH < Amos wash, long incubation, AHG Positive for T-lymphocytes : Ab against HLA class I (A, B, C) Close association with hyperacute rejection → Absoulte contra-indication for transplantation Positive for B-lymphocytes : Ab against HLA class II (DR, DQ), weak class I, non. HLA Ag (ex. auto. Ab)