固相生物素－亲和素ELISA检测HIV抗体方法的建立与应用

The Establishment and Application Of Solid Phase Biotin Avidin ELISA Determinating HIV Antibodies

采用生物素化ＨＩＶ合成肽抗原与预吸附在ＥＩＡ反应板上的亲和素结合，建立检测ＨＩＶ抗体的固相生物素－亲和素ＥＬＩＳＡ方法。经ＨＩＶ（１＋２）抗体参考血清Ｐａｎｅｌ检定，其灵敏度（１３／１３）１００％，特异性（２８／２８）１００％，批内变异系数＜７％，批间变异系数＜１０％。１０５６８例确认血清标本验证，阳性检出率１００％，阴性总符合率９９．８％，假阳性率０．２％。与常规间接ＥＬＩＳＡ法比较，本法操作简便，试验所需时间短，灵敏度。

The establishment of the sloid phase Biotin avidin ELISA method to determinate HIV(1+2) antibodies involves the utilization of biotinazed synthetic HIV peptides gp120,gp36,gp24,gp41 combined with the avidin coated on EIA plates, and anti human IgG McAb HRP. The result of HIV(1+2)antibody reference serum panels established by National Institute for the Control o Pharmacentical and Biological Products showed that the sensitivity was 13/13,Speciticity was 28/28,the coefficient of variations(cv) were below 10% intra assay and inter assay;44 HIV antibody positive serum samples confirmed by western blotting test using Bio Rad(USA) regents were positive by our Kit test, the coincidence rate was 99.8% in 10537 HIV antibody nagative serum samples, the false positive reaction rate was 0.2%. The method increases the density of effective angigen coated on the EIA reaction plate and overcomes the disadvantage of weak absorption and rapid degradation of small peptides due to the direct coating to EIA reaction plate. so the method significantly improve the sensitivity, speciticity and stability in the determination of HIV antibody. The method is also rapid, simple and inexpensive. it is better ELISA technique for detecting of HIV antibodies.