Establishment and preliminary verification of indirect ELISA for detection of serum antibody against varicella-zoster virus
10.3760/cma.j.cn112309-20241210-00420
- VernacularTitle:水痘-带状疱疹病毒血清抗体检测间接ELISA法的建立及初步验证
- Author:
Wei LI
1
;
Xinyi WANG
1
;
Jilai LI
1
;
Duoqian WEI
1
;
Shuxiang LI
1
Author Information
1. 国药中生生物技术研究院（新型疫苗国家工程研究中心）第三研究室，北京 101111
- Publication Type:Journal Article
- Keywords:
Varicella-zoster virus;
Indirect ELISA;
Antibody;
Fluorescent antibody to membrane antigen (FAMA) test
- From:
Chinese Journal of Microbiology and Immunology
2025;45(9):783-787
- CountryChina
- Language:Chinese
-
Abstract:
Objective:To establish and verify an indirect ELISA for quantitative detection of serum antibody against varicella-zoster virus (VZV).Methods:Recombinant VZV glycoprotein E (gE) was diluted with the coating buffer to concentrations of 2, 0.5, and 0.25 μg/ml. HRP-labeled antibody dilutions were 1∶2 000, 1∶5 000, and 1∶10 000. The optimal coating concentration and the optimal working concentration were determined according to the A450/630 value and the P/N ratio (working reference A450/630 value/blank control A450/630 value). According to the indirect ELISA process, an indirect ELISA quantitative detection method for VZV-specific antibody was established. The linear range, accuracy, and precision of the method were verified. The titers of VZV-specific IgG antibody in 32 serum samples were detected by the established method, fluorescent antibody to membrane antigen (FAMA) test, and commercially available VZV quantitative ELISA kit, and the results of the three methods were compared. Results:The optimal coating concentration was 0.25 μg/ml and the optimal working concentration of HRP-labeled antibody was 1∶10 000. The linear range of the standard curve was 0.015 7-1 mIU/ml; the recovery rate was 95.32%-110.99%, and the coefficient of variation (CV) values for repeatability and the intermediate precision were both less than 10%. The results of the established indirect ELISA were well correlated with the results of FAMA test and commercial ELISA kit (indirect ELISA & FAMA: the correlation coefficient was 0.886 4, indirect ELISA & commercially ELISA kit: the correlation coefficient was 0.806 6).Conclusions:The established indirect ELISA method for serum anti-VZV antibody detection has good accuracy and precision, and shows a good correlation with FAMA test and commercial VZV ELISA kit.
