液相芯片法验证蛋白质免疫印迹试验检测HIV-1结果的敏感性和特异性

Sensitivity and specificity analyses on HIV-1 results detected by Western Blot method using liquid chip technology

目的初步分析采用液相芯片法研制人类免疫缺陷病毒(HIV)-1检测试剂的可行性。方法采用液相芯片技术用蛋白质免疫印迹试验(Western Blot)检测HIV-1阳性和阴性的标本;用HIV-1的gp120、gp41、p24抗原分别包被不同编号的免疫磁珠,与生物素化二抗、亲和素化荧光染料PE组成液相芯片检测系统,对标本进行检测分析。结果 55份标本经Western Blot法确认阳性样本检测符合率为100%;76份经酶联免疫吸附试验(ELISA)初检和复检阴性的标本检测HIV-1gp120、HIV-1gp41、HIV-1p24抗原均为阴性;86份经ELISA法检测初检和复检为阳性而Western Blot法确认为阴性的样本,其HIV-1gp120、HIV-1gp41、HIV-1p24单片段检测均为阴性。液相芯片法检测HIV-1抗体的敏感度和特异度均为100%。结论采用液相芯片技术组成的HIV-1抗体检测系统,其敏感度和特异度均优于ELISA法检测。

Objective Preliminary analysis of the feasibility of using liquid chip method to develop human immunodeficiency virus（HIV）-1 detection reagent. Methods Western Blot method with liquid chip technology was applied to detect the confirmed HIV-1 positive and negative samples. The liquid chip detection system was formed by different code numbers of immunomagnetic beads coated with HIV-1 gp120, gp41, p24 antigens respectively, with biotinylated second antibody and avidin fluorescent dye PE to detect and analyze the variety of samples. Results The coincidence rate was 100% based on the analysis of 55 HIV-1 positive samples confirmed by Western blot; after examination and reexamination of 76 samples by enzyme linked immune sorbent assay（ELISA）, the HIV-1 results were all negative, and the detection of such samples for HIV-1 gp120, HIV-1 gp41, and HIV-1 p24 antigens were also all negative; after ELISA examination and reexamination for 86 samples, the HIV-1 results were positive, but after using Western Blot detection for such samples, the results were definitely negative, and the detections of single fragments of HIV-1 gp120, HIV-1 gp41, HIV-1 p24 were also all negative. Conclusion The sensitivity and specificity of liquid chip technology used in Western Blot method for HIV-1 antibody detection system are superior to those in ELISA detection.