Comparison of Multiple Passage Integrated Cell Culture-PCR and Cytopathogenic Effects in Cell Culture for the Assessment of Poliovirus Survival in Water |
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Authors: | Madhumita Mahalanabis Kelly A Reynolds Ian L Pepper Charles P Gerba |
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Institution: | (1) Department of Biomedical Engineering, Boston University, 44 Cummington Street, Boston, MA 02215, USA;(2) Environmental Health Sciences, Mel & Enid Zuckerman College of Public Health, The University of Arizona, 1295 N. Martin, 245163, Tucson, AZ 85724, USA;(3) Department of Soil, Water, and Environmental Science, The University of Arizona, 1200 E. South Campus Drive, 429 Shantz Building #38, P.O. Box 210038, Tucson, AZ 85721, USA; |
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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. |
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