基于HSV-1型特异性表位的血清抗体ELISA检测方法建立与初步应用评价

An ELISA based serum HSV-1 specific epitope antibody determination: methodology and preliminary application

目的建立并评价HSV-1型特异性抗体鉴别诊断的ELISA方法。方法以HSV-1-gG112-127型特异性表位的串联重组表达蛋白作为包被抗原,建立检测血清HSV-1特异性IgG的间接ELISA方法。同时以免疫印迹检测作为“金标准”,评价检测方法的真实性和可靠性。结果采用棋盘法确定了包被抗原、抗体的最佳浓度,建立ELISA检测方法。血清特异性IgG检测的灵敏度为91%,特异度为97.6%,阳性预测值为97%,阴性预测值为93%,符合率为94.5%,试验的一致率为98%。结论用串联重组蛋白作为包被抗原对HSV-1感染的血清进行ELISA分型检测,其特异性好。

Objective To establish and evaluate an EHSA method for determining the serum specific antibody against HSV-1. Methods A recombinant protein based on the type-specific epitope of HSV- 1 glycoprotein G112-127 was used as coated antigen to establish a diagnositic method of ELISA for detecting the specific IgG in sera, and Western blot technique was considered as the ＂golden standard＂ to evaluate its specificity and reliability. Results After chess titration of the HSV-1 antigen and the secondary antibody, an indirect EHSA was established. The sensitivity, specificity, positive predictive value, negative predictive value, coincidence rate, concordance rate of the constructed method were 91%, 97.6%, 97%, 93 %, 94.5 %, and 97 %, respectively. Conclusion The recombinant protein could be used as coat antigen to detect the specific antibody against HSV-1.