IgA 缺乏症献血者酶联免疫试验筛查方法的建立

摘要

目的：建立适合用于大规模IgA缺乏症献血者筛选的ELISA检测方法。方法采用间接酶联免疫法，在微孔板中包被羊抗人IgA、用辣根过氧化物酶标记羊抗人IgA抗体作为酶标二抗。结果建立的ELISA测定法灵敏度为0．1μg／mL ，IgA浓度为0．1、100μg／mL的批间变异系数（CV ）是1．74％、3．49％，批间CV中位数为3．48％（范围：1．83％～6．96％）。检测过程约80 min。结论成功建立了IgA缺乏症筛查的ELISA测定法，其灵敏度高、特异度强、省时、操作简易，可用于大规模筛选IgA缺乏症献血者及建立Ig A缺乏献血者库。%Objective To establish the enzyme‐linked immuno sorbent assay(ELISA) method for a large‐scale screening of IgA deficiency in blood donors .Methods The indirect ELISA was adopted .The goat anti‐human IgA antibody was coated in microwell plates and labeled by horse radish peroxidase (HRP) as enzyme‐labelled secondary antibody .Results The sensitivity of established ELISA detection method was 0 .1 μg/mL .The intraassay coefficients of variation (CV) for IgA concentrations of 0 .1 ,100 μg/mL were 1 .74% to 3 .49% .The median interassay CV was 3 .48% (range:1 .83% -6 .96% ) .The assay process was 80 min . Conclusion TheELISA detection method is successfully established with high sensitivity ,strong specificity ,timesaving and easy operating and can be used for a large scale screening of Ig A deficiency and establishment of blood donors bank of Ig A deficiency .