Comparison of Multiple Passage Integrated Cell Culture-PCR and Cytopathogenic Effects in Cell Culture for the Assessment of Poliovirus Survival in Water

Abstract

The goal of this study was to determine if a cytopathogenic effects (CPE) cell culture assay and an integrated cell culture PCR (ICC-PCR) assay would yield similar or different results when used to assess virus survival in water. Poliovirus type 1 was added to dechlorinated tapwater and stored at room temperature (22.5–24°C) for a total of 50 days. Samples were assayed at defined time intervals by the most probable number (MPN) method on Buffalo green monkey kidney cells (BGM) by CPE and additionally by ICC-PCR. Monolayers that were CPE negative on first passage were passed onto fresh monolayers of cells for a second and third time if still negative. By CPE assay, second passage was observed to yield a greater titer (2,300 vs. 24,000 MPN/ml) and third passage also resulted in an increased titer. ICC-PCR proved to be a more rapid and sensitive method than conventional cell culture for determining virus inactivation rates in water. Poliovirus survived in tapwater for up to 32 days, as assessed by both third passage ICC-PCR and CPE. There was no statistical difference in the inactivation rates between the two methods. To determine the total number of infectious viruses, these findings indicate the need for performing three cell culture passages or, alternatively, ICC-PCR on first passage.