Validation of a cell infection-based quantitative RT-PCR for evaluation of rotavirus vaccine potency

Abstract

Objective To validate a cell infection-based quantitative RT-PCR for evaluating the potency of rotavirus vaccine. Methods According to the ICH (the International Council for Harmonization) Harmonised Tripartite Guideline, the method was validated for its specificity, accuracy, precision, linearity and robustness. Results The 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. Conclusions The 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