水痘-带状疱疹病毒膜抗原荧光抗体试验的建立与评价

摘要

目的 建立用于检测血清中水痘-带状疱疹病毒(varicella-zoster viruS,VZV)特异性抗体的膜抗原荧光抗体(fluorescent antibody to membrane antigen,FAMA)试验并对其进行验证和评价.方法 用VZV感染细胞作为抗原制备抗原载玻片,以异硫氰酸荧光素标记的羊抗人IgG为二抗建立 FAMA试验.对试验的灵敏度、特异性和重复性进行验证.用FAMA试验和市售ELISA试剂盒对200份血浆样品中的抗VZV抗体进行检测.采用Spearman秩相关检验对两种方法的检测结果进行比较.结果 FAMA试验的灵敏度为0.04 IU/ml,与常见的人疱疹病毒(单纯疱疹病毒1和2型、人巨细胞病毒)没有交叉反应且重复性良好.经FAMA试验检测,200份血浆样品的抗体阳性率为93.5％,抗体几何平均效价为1 : 18.1,检测结果与市售ELISA试剂盒的符合率为90. 0％.两种方法的检测结果具有相关性,Spearman秩相关系数为0.49,P <0.000 1.结论 建立的方法具有良好的灵敏度、特异性和重复性,可用于血清中抗VZV抗体的检测.%Objective To develop and evaluate a fluorescent antibody to membrane antigen(FAMA) test for detection of varicella-zoster virus(VZV)-specific antibody in plasma. Methods The FAMA test was established with VZV-infected cells as fixed antigen and fluorescein isothiocyanate labeled goat anti-human IgG as secondary antibody. The sensitivity, specificity and repeatability of the technique were validated. The antibodies in 200 plasma samples were determined with the established test and a commercial ELISA kit. The detection results of two methods were compared using Spearman's rank correlation test. Results FAMA test had high sensitivity with the detecting limit of 0.04 IU/ml. No cross reaction with other herpesviruses, such as herpes simplex virus (HSV) 1, 2 and human cytomegalovirus was observed. The antibody positive rate and geometric mean titer of 200 samples determined by FAMA test were 93.5％ and 1: 18.1, respectively. The result coincidence rate of FAMA test with the ELISA kit was 90.0％. The results of two methods were correlated with Spearman's rank correlation coefficient 0.49, P <0.000 1. Conclusion The developed method has good sensitivity, specificity and repeatability, and can be applied for detecting anti-VZV antibodies in sera.