Test Technology
Validation of a cell infection-based quantitative RT-PCR for evaluation of rotavirus vaccine potency
Yueyue Liu, Yunqi Zhang, Yan Liu, Yunjin Wang, Mingqiang Wang, Yan Zhao, Jialiang Du, Chao Ma, Xu Zhou, Tai Guo
Published 2019-07-31
Cite as Chin J Microbiol Immunol, 2019, 39(7): 532-537. DOI: 10.3760/cma.j.issn.0254-5101.2019.07.008
Abstract
ObjectiveTo validate a cell infection-based quantitative RT-PCR for evaluating the potency of rotavirus vaccine.
MethodsAccording to the ICH (the International Council for Harmonization) Harmonised Tripartite Guideline, the method was validated for its specificity, accuracy, precision, linearity and robustness.
ResultsThe method had good specificity as it could only amplify and detect the corresponding type of rotavirus strain. The recovery rates for determining the potency against rotaviruses of G2, G3 and G4 types were 97% to 108%. The percent coefficient of variation (CV) of both intra-plate and inter-plate precision was ≤2.62%, while the intraday and interday CV was ≤1.76% and ≤2.27%, respectively. The CV between the two experimenters was ≤7.68%. The linearity range of the method was 4.4-6.5 UI for G2 type rotavirus, 3.9-8.3 UI for G3 type and 3.5-8.1 UI for G4 type. Good robustness was observed using the cells of 140 to 160 generations.
ConclusionsThe cell infection-based quantitative RT-PCR was shown to have satisfactory specificity, accuracy, precision, linearity and robustness, suggesting that it was a suitable method for evaluating the potency of multivalent rotavirus live vaccines.
Key words:
Rotavirus; Vaccine; Potency assay; Validation
Contributor Information
Yueyue Liu
Department of Enteric Virus Vaccine, National Institutes for Food and Drug Control, Beijing 102629, China
Yunqi Zhang
School of Mathematics and Statistics, Yunnan University, Kunming 650091, China
Yan Liu
Department of Enteric Virus Vaccine, National Institutes for Food and Drug Control, Beijing 102629, China
Yunjin Wang
The Second Laboratory, Lanzhou Institute of Biological Products Co. Ltd., Lanzhou 730046, China
Mingqiang Wang
The Second Laboratory, Lanzhou Institute of Biological Products Co. Ltd., Lanzhou 730046, China
Yan Zhao
Department of Enteric Virus Vaccine, National Institutes for Food and Drug Control, Beijing 102629, China
Jialiang Du
Department of Enteric Virus Vaccine, National Institutes for Food and Drug Control, Beijing 102629, China
Chao Ma
The Second Laboratory, Lanzhou Institute of Biological Products Co. Ltd., Lanzhou 730046, China
Xu Zhou
Shanghai Institute of Biological Products Co. Ltd., Shanghai 200052, China
Tai Guo
Department of Enteric Virus Vaccine, National Institutes for Food and Drug Control, Beijing 102629, China
