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目的评价检测艾滋病病毒Ⅰ型(HIV-1)新发感染的限制性抗原亲和力酶联免疫试验的重复性、稳定性和准确度,探讨该方法推广应用的可行性。方法采用检测精密度、孔间精密度、批间差、两实验员和两实验室之间结果的相关性,评价试剂的重复性;利用37℃热稳定性试验评价试剂的稳定性;使用国际标准盘评价试剂的准确度。结果检测精密度结果显示,所有样本标化吸光度值(ODn)的变异系数(CV)<15%,两实验员和两实验室试验结果的相关系数R2值均>0.99,孔间精密度CV值最小为3.76%,3个批次试剂的批间差结果显示,25份内控盘样本中,只有1份样本CV值>15%,为22.5%。37℃放置7天与4℃保存的酶结合物和酶联板,对内控盘样本检测的ODn值没有明显区别。国际标准盘评判结果与预期值一致。结论检测HIV-1新发感染的限制性抗原亲和力方法具有较好的稳定性、重复性和准确度,可以在实验室网络中推广应用。
Objective To evaluate the repeatability, stability and accuracy of restriction enzyme immunoassay (ELISA) for the detection of HIV-1 infection and to explore the feasibility of its application. Methods The repeatability of the reagents was evaluated by the correlation between the precision of the test, the precision of the wells, the difference between batches, the results of the two laboratory workers and the two laboratories. The stability of the reagents was evaluated by the thermal stability test at 37 ℃. The standard plate is used to evaluate the accuracy of the reagents. Results The test results showed that the coefficient of variation (CV) of the normalized absorbance (ODn) of all samples was less than 15%, the correlation coefficient R2 between two laboratory technicians and the two laboratory test results was> 0.99, and the CV The minimum batch was 3.76%. The batch-to-batch difference between the three batches of reagents showed that only 1 out of 25 samples of control disks had CV> 15%, which was 22.5%. 37 ℃ placed 7 days and 4 ℃ stored enzyme conjugate and ELISA plate, the ODn value of the inner control plate sample no significant difference. The result of the international standard plate is consistent with the expected value. Conclusion The method of detecting the affinity of restricted antigen of HIV-1 new infection has good stability, repeatability and accuracy and can be widely applied in the laboratory network.