HIV-1/2抗体和P24抗原联合检测方法的建立及应用

目的建立HIV-1/2抗体和P24抗原联合检测的酶免疫检测方法,用于HIV感染的实验室诊断,缩短HIV感染的检测窗口期。方法制备抗HIVP24单克隆抗体和多克隆抗体,联合使用基因工程HIV1/2型抗原和抗HIVP24单克隆抗体包被酶联反应板,以辣根过氧化物酶标记的HIV1/2型抗原和生物素化的兔抗HIVP24抗体作为标记物,建立联合检测HIV1/2抗体和P24抗原的ELISA方法,并对其特异性、敏感性和稳定性进行鉴定。结果检测P24抗原质控品的灵敏度可达0.2ng/ml;检测187份HIV阳性血清、210份其他疾病患者血清和520份正常人血清,其敏感性和特异度分别为100%和99.62%。同时重复12次检测4份HIV阳性血清,变异系数均小于10%。结论本方法可以在1.5h内一步同时检出特异性HIV-1/2抗体和HIVP24抗原,缩短了HIV感染的检测窗口期,具备快速、简便、特异、敏感等优点,适用于HIV感染的实验室诊断和流行病学调查。

Establishment and Application of an Enzyme Immunoassay Combined With Anti - HIV1/2 Antibody and P24 Antigen

Objective To establish an enzyme immunoassay combined with anti-HIV1/2 antibody and P24 antigen for shortening the examination window period of HIV infection in HIV laboratory diagnosis.Method Anti-HIV P24 monoclonal and polyclonal antibodies were prepared.Applying HIV1/2 antigen and anti-HIV P24 antibody coated enzyme-linked reaction plates,labeling HIV1/2 antigen and anti-HIV P24 antibody with horseradish peroxidase,and setting up an integrated enzyme-linked Immunosorbent assay(ELISA)for the test of anti-HIV-1/2 antibody and HIV P24 antigen,the specificity and sensitivity of this method were tested and validified.Results The sensitivity of P24 antigen control test was up to 0.2 ng/ml.A total of 187 serum samples of patients with AIDS,210 serum samples of patients with other diseases and 520 serum samples of healthy subjects were examined and compared,and the results revealed the sensitivity was 100% and the specificity was 99.62%.Conclusion The anti-HIV1/2 antibody test and the HIV P24 antigen test can be measured at the same time within 1.5 hours by this rapid,simple,specific and sensitive method,while the examination window period of HIV infection is shortened.Thus,the method is suitable for laboratory diagnosis and epidemiological investigation.