Validation of a pseudovirus-based neutralization assay for detection of human papillomavirus antibodies in human serum samples / 中华微生物学和免疫学杂志

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.