合成肽抗原抗人免疫缺陷病毒1/2型抗体酶联试剂盒的研制

The detection of human immunodeficiency virus type 1/2 antibody using synthetic peptides

根据人免疫缺陷病毒（ＨＩＶ）的基因结构和氨基酸序列，采用固相法合成了ＨＩＶ－１ｇｐ４１（ＳＰ１）、ｇｐ１２０（ＳＰ２）、ｐ２４（ＳＰ３）和ＨＩＶ－２ｇｐ３６（ＳＰ４）的４条多肽，混合包被酶标板做为固相抗原，采用间接酶联免疫吸附试验（ＥＬＩＳＡ），建立了检测抗－ＨＩＶ－１／２ＩｇＧ抗体的酶联诊断试剂盒。检测卫生部药品和生物制品检定所提供的４１份质控参比血清，其特异性、敏感性均为１００％，变异系数小于１０％。检测１８６份其它病种病人血清均为阴性，与华怡、巴斯德、金豪等公司的ＨＩＶ诊断试剂比较检测了９０份ＨＩＶ感染者和１４０份正常人血清，除与华怡试剂的阴、阳性及总符合率分别为９９２９％（１３９／１４０）、９８９０％（９０／９１）和９９５７％（２２９／２３０）外，其余均为１００％。３７℃放置４天后试剂的检测结果不受影响。

Four peptides of human immunodeficiency virus (HIV) located in the regions of gp41, gp120, p24 of HIV-1 and gp36 of HIV-2 have been synthesized according to the published amino acid sequences and the positions of antigenetic determinants of viruses by a solid-phase method. An indirect ELISA for detecting antibody against HIV 1/2 was established using these synthetic peptides as the coating antigens. We detected 41 control sera provided by National Institute for the Control of Pharmaceutical and Biological Products. The results indicated that the general coincident rate were 100%, the variant coefficient was less than 10%. A comparison of our reagent with Huayi, Pasteur and Jinhao reagents in detection of 90 positive sera and 140 normal sera for HIV -1/2 showed that the coincident rate was 100% with Pasteur and Jinhao, 99.57% with Huayi. All of 186 sera from patients with other diseases were negative. The reagent are stable at 37℃ for 4 days. The results indicated that our reagent is highly specific, sensitive and stable, it can be used for detection of HIV -1/2 infections.