以多肽为抗原的 HIV ELISA 检测试剂的研究与开发

摘要

目的：采用四种特异性化学合成多肽抗原（ HIV-Ⅰ gp41、gp120、p24和HIV-Ⅱ gp36）作为包被主要原材料，制备成检测HIV-Ⅰ/Ⅱ抗体的试剂盒。方法用棋盘滴定法选择合适的抗原包被浓度及酶标抗原的工作浓度，经过优化检测系统后，对本试剂盒进行相关的性能验证，并以上海科华、厦门英创及珠海丽珠试剂作为对照。结果研究得出合适包被抗原浓度为5μg/ml，酶标抗原工作浓度为1∶160。经过优化检测系统试剂的检测，CUT-OFF值为0.207，灵敏度为97.2％，特异性为98.6％。试剂主要活性成分的稳定性及检测的重复性良好，与对照试剂检测结果一致。结论以化学合成多肽抗原为主要包被原材料制备成检测HIV抗体的试剂盒具有可行性，试剂盒主要活性成分稳定，且具有较好的检测敏感度和特异性。%Objective To prepare diagnostic kit for HIV-Ⅰ/Ⅱ antibodies with mixed polypeptide antigen including HIV-Ⅰgp41, gp120, p24 and HIV-Ⅱ gp36.Methods The appropriate concentration of coated antigen and HRP-Ag were selected by checkerboard titration, and the sensitivity and specificity of the kit were conducted.Re-sults The appropriate concentration of coated antigen was 5 μg/ml, and the appropriate concentration of HRP-Ag was 1∶160.After optimizing the detection system, the CUT-OFF value, sensitivity and specificity of the kit were 0.207, 97.2%and 98.6%, respectively.Conclusion It is feasible that detect HIV infection via polypeptide anti-gen coated in polystyrene plate with high sensitivity and specificity.