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目的优化蛋白免疫印迹法(WB)对艾滋病病毒(HIV)抗体不确定血清的检测条件,并测试其检测效能。方法采用正交实验L9(34)法,对HIV弱阳性对照的WB检测条件(样本量、样本孵育时间、酶联合物孵育时间)进行优化设计;对19份HIV抗体不确定血清、25份HIV阴性血清和20份HIV阳性血清,同时采用标准检测条件和优化检测条件进行检测。结果以样本量100μL、样本孵育时间2小时、酶联合物孵育时间15分钟,为HIV抗体不确定血清的最优检测条件;同标准检测条件相比,优化检测条件对HIV阴性或阳性血清的检测效能一致,但对19例HIV抗体不确定血清的检测效能更高。结论正交实验所筛选的优化检测条件可高效检测HIV抗体不确定血清。
OBJECTIVE: To optimize the detection conditions of uncertain serum of HIV antibody by Western blotting (WB) and test its detection efficiency. Methods The orthogonal design of L9 (34) method was used to optimize the WB detection conditions (sample size, sample incubation time and enzyme conjugate incubation time) of HIV weakly positive control. Nineteen HIV positive serums, 25 HIV Negative sera and 20 HIV-positive sera were tested simultaneously using standard and optimized assay conditions. Results The optimum conditions for the determination of HIV antibody undefined serum were 100 μL sample size, 2 hours sample incubation time and 15 minutes incubation time. Compared with the standard test conditions, the detection conditions of HIV negative or positive serum were optimized Efficacy, but 19 cases of HIV antibody detection of undefined serum higher efficiency. Conclusion The optimal test conditions screened by orthogonal experiment can detect HIV antibody uncertain serum efficiently.