Validation of a pseudovirus-based neutralization assay for detection of human papillomavirus antibodies in human serum samples
10.3760/cma.j.issn.0254-5101.2018.07.010
- VernacularTitle: 人乳头瘤病毒假病毒中和抗体检测方法在临床样本检测中的验证
- Author:
Jianhui NIE
1
;
Tingting NING
;
Ruifeng CHEN
;
Qing WANG
;
Weijin HUANG
Author Information
1. Division of HIV/AIDS and Sex-transmitted Virus Vaccines, National Institutes for Food and Drug Control, Beijing 102629, China
- Publication Type:Journal Article
- Keywords:
Human papillomavirus;
Neutralizing antibody;
Validation
- From:
Chinese Journal of Microbiology and Immunology
2018;38(7):529-534
- CountryChina
- Language:Chinese
-
Abstract:
Objective:To validate a pseudovirus-based neutralization assay for the detection of antibodies against human papillomavirus (HPV) in human serum samples.
Methods:The specificity, accuracy, precision, range of linearity, limit of detection and robustness of the neutralization assay were evaluated using HPV-negative serum samples, vaccinated serum samples with quadri-valent vaccine, and international standards for detecting antibodies against HPV16 and HPV18.
Results:Based on the data of the HPV-naïve samples, the criteria of positivity was determined as follows: the 50% inhibitory dose (ID50) of the tested sample was not less than 40 and 2-fold not less than that for bovine papillomavirus. The neutralization assay showed good accuracy with a recovery rate of 87%-122% and excellent reproducibility with intra- and inter-assay variation of 5%-27% and 10%-26% respectively. The HPV16 and HPV18 international standards were used to define the limit of quantification, which was 1.28 IU/ml for HPV16 and 0.96 IU/ml for HPV18. Acceptable ranges of variation for the key parameters of this assay were defined, which showed the good robustness of the pseudovirus-based neutralization assay.
Conclusion:The pseudovirus-based neutralization assay for the detection of HPV antibodies showed good specificity, accuracy, sensitivity, and robustness, suggesting that it could be used to evaluate the immunogenicity of HPV vaccines.
