TP-ELISA试剂盒3种外膜脂蛋白抗原联合应用对检测结果影响分析

Analysis of the effect of three kinds of outer membrane lipoprotein antigens combined with TP-ELISA on the detection results

目的通过对大连地区无偿献血者梅毒单试剂阳性和双试剂阳性血清标本中TP15、TP17、TP47抗体分别进行检测,分析梅毒A试剂所包被的这3种不同外膜脂蛋白抗原对检测结果的影响。方法收集大连市血液中心2017年11月—2018年11月梅毒A试剂单阳性标本36份和171份TP双试剂阳性标本,采用A试剂厂家3种分别包被不同外膜脂蛋白抗原的TP-ELISA试剂进行检测,所有阳性标本都进行电化学发光和TPPA确证实验,记录结果,并对数据进行统计分析。结果 1)TP单试剂阳性(A厂家)和双试剂阳性标本进行确认实验后,假阳性率分别为100%和10.83%(P<0.05)。2)单试剂阳性(A厂家)标本36份,3种抗体检测结果筛查频率P分别为:TP15-,TP47-,TP17-(P=0.17);TP15+,TP47-,TP17-(P=0.22);TP15-,TP47+,TP17-(P=0.56);TP15-,TP47-,TP17+(P=0.06);TP15+,TP47+,TP17-、TP15-,TP47+,TP17+、TP15+,TP47-,TP17+、TP15+,TP47+,TP17+频率P均为0。3)梅毒双试剂阳性标本(确认阳性)140份,3种抗体检测结果筛查频率P分别为:TP15+,TP47-,TP17-(P=0.01);TP15-,TP47+,TP17-(P=0.01);TP15+,TP47+,TP17-(P=0.04);TP15-,TP47+,TP17+(P=0.01);TP15+,TP47-,TP17+(P=0.01);TP15+,TP47+,TP17+(P=0.92);TP15-,TP47-,TP17-和TP15-,TP47-,TP17+频率P为0。4)梅毒单阳性的标本3种抗体检测结果两两进行χ~2配对检验,TP17和TP47 2种抗体的阳性检出率有差异(P<0.05);TP17和TP15 2种抗体的阳性检出率无差异(P>0.05);TP15和TP47 2种抗体的阳性检出率有差异(P<0.05)。5)梅毒双试剂阳性的标本(确认阳性)3种抗体检测结果两两进行χ^2配对检验均无差异(P>0.05)。结论 TP-ELISA检测试剂A包被的TP47外膜脂蛋白可能是引起其假阳性率升高的主要原因,针对TP47外膜脂蛋白在梅毒检测试剂A中的应用价值试剂厂家应该进行进一步的研究。

Objective To analyze the influence of three kinds of outer membrane lipoprotein antigens coated with Syphilis A reagent on the detection results by testing the antibodies of TP15, TP17 and TP47 in the serum samples of voluntary blood donors with reactivity in single-and dual-reagents for Syphilis in Dalian area.Methods Thirty-six samples with single-reactivity in Syphilis A reagent and 171 with dual-reactivity were collected from Dalian Blood Center from November 2017 to November 2018. These samples were detected by three TP-ELISA reagents coated with different foreign model lipoprotein antigens from A reagent manufacturer. All positive specimens were confirmed by electrochemical luminescence and TPPA. The results were recorded, and the data were statistically analyzed.Results 1)After the confirmation test of single-and dual-reactive TP reagents(manufacturer A), the false positive rates were 100% and 10.83%(P<0.05). 2) There were 36 samples active with single reagent(manufacturer A), and the screening frequencies of these three antibody detection results were: TP15-,TP47-,TP17-(P=0.17);TP15+,TP47-,TP17-(P=0.22);TP15-,TP47+,TP17-(P=0.56);TP15-,TP47-,TP17+(P=0.06);TP15+,TP47+,TP17-(P=0),TP15-,TP47+,TP17+(P=0), TP15+,TP47-,TP17+(P=0) and TP15+,TP47+,TP17+(P=0). 3)There were140 Syphilis confirmed with dual TP reagents, and the screening frequency(P) of the three antibody test results were: TP15+,TP47-,TP17-(P=0.01);TP15-,TP47+,TP17-(P=0.01);TP15+,TP47+,TP17-(P=0.04);TP15-,TP47+,TP17+(P=0.01);TP15+,TP47-,TP17+(P=0.01);TP15+,TP47+,TP17+(P=0.92);TP15-,TP47-,TP17-and TP15-,TP47-,TP17(P=0). 4)The three antibody test results of single-positive Syphilis samples were paired with χ~2 test, and there was significant statistical difference in the positive detection rates of TP17 vs. TP47 and TP15 vs. TP47(P<0.05), while no statistical difference in TP17 vs. TP15(P>0.05). 5) No significant difference was observed as the test results of the three antibody samples confirmed by dual-reagents were compared in pairs by χ~2 pairing test(P>0.05).Conclusion TP47 outer membrane lipoprotein coated with TP-ELISA reagent A may be the main cause of false positive rate. Moreover, there was no significant difference between TP47 and TP15/TP17 in the detection of true positive Syphilis. Therefore, it suggests that the application of TP47 outer membrane lipoprotein in Syphilis detection reagent A should be further weighed by the reagents manufacturers.