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毒物分析 免疫檢驗. 賴滄海教授 03-07-2008. Characteristics of the Screening and Confirmation Assays. Screening Assays Easy to Perform (automation) Low cost Sensitivity Precision Accuracy Confirmation Assays Accuracy Precision Sensitivity Low Cost Easy to Perform. 濫用藥物免疫檢驗試劑之原理 ( 一 ).
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毒物分析 免疫檢驗 賴滄海教授 03-07-2008
Characteristics of the Screening and Confirmation Assays • Screening Assays • Easy to Perform (automation) • Low cost • Sensitivity • Precision • Accuracy • Confirmation Assays • Accuracy • Precision • Sensitivity • Low Cost • Easy to Perform
濫用藥物免疫檢驗試劑之原理 (一) 1. Homogenous Immunoassay • Enzyme immunoassay Enzyme Multiplied Immunoassay Technique (Syva EMIT), DRI. Cloned Enzyme Donor Immuno Assay (CEDIA). • Fluorescence immunoassay Fluorescence Polarization Immunoassay (FPIA). Abbot TDx. • Microparticle agglutination immunoassay Roche Abuscreen Online
濫用藥物免疫檢驗試劑之原理 (二) 2. Heterogeneous Immunoassay • Radio immunoassay (Roche, DPC) • Enzyme Linked Immunoassay (DCB)
G6PD -* + Ab → G6PD -*:Ab (↓Enzyme activity) + * in sample G6PD -* (Full Enzyme activity) + G6PD-*:Ab (↓Enzyme activity) + *:Ab G6PD -* G6P + NAD+6-phosphogluconate + NADH (↑A 340) EMIT的原理
EA + ED* + Ab →Ab :*+ (EA-ED*)4 Active enzyme –* ED*: Ab + EA No enzyme activity Schematic Illustration of CEDIA
Ab + Ag - Fl → Ab:Ag -Fl (Polarized Light) Ag Ab:Ag + Ab:Ag -Fl + Ag - Fl (Decreased Polarization) Fluorescence Polarization Immunoassay (FPIA)
DRUG * DRUG + Ab1 DRUG * DRUG DRUG * :Ab1 DRUG :Ab1 + Ab2 DRUG *:Ab1 :Ab2 DRUG :Ab1 :Ab2 Schematic Illustration of RIA (ppt)
-* + substrate Plate -Ab + *-Enz wash Enzyme Activity +* wash + substrate No enzyme activity Schematic illustration of DCB EIA
Percent cross reactivity of amphetamine analogs with various immunoassays (1)
Percent cross reactivity of amphetamine analogs with various immunoassays (2)
Percent cross reactivity of amphetamine analogs with various immunoassays (3)
Percent cross reactivity of amphetamine analogs with various immunoassays (4)
Percent cross reactivity of amphetamine analogs with various immunoassays (5)
Apparent cross-reactivity (%) of various opiate immunoassays
Adulterants that Affects the Immunoassays 1. In Vivo Adulteration. 2. In Vitro Adulteration. (1) Detecting Adulteration. (2) EIA are more likely to be affected. (3) Substances used as adulterants. Alcohols, Ammonia, Ascorbic acid, Bleach, Blood, Detergent, Drano, Lemon Juice, Peroxide, Salt, Soap, Visine, Urinaid.
C:品管線(陽性反應線) T:藥物結合線(陰性反應線) 水分吸收區 C T 反應區: 抗體-色素結合物 樣品吸收區 C C T 陽性 陰性 單一步驟免疫層析法 (One-Step Immunochromatography ) (二條藍色線條) (一條藍色C線)