Development and Validation of an Integrated Cell Culture-qRTPCR Assay for Simultaneous Quantification of Coxsackieviruses, Echoviruses, and Polioviruses in Disinfection Studies

Document Type

Article

Language

eng

Format of Original

13 p.

Publication Date

2010

Publisher

IWA Publishing

Source Publication

Water Science & Technology

Source ISSN

0273-1223

Original Item ID

doi: 10.2166/wst.2010.818

Abstract

This study demonstrated the applicability of integrated cell culture-quantitative RTPCR (ICC-qRTPCR) for the simultaneous quantification of coxsackievirus, echovirus, and poliovirus in disinfection studies. Buffalo green monkey cells were inoculated with a 10-fold dilution series of mixed enteroviruses and incubated prior to qRTPCR quantification. Optimal assay conditions included three post infection washes and a 24-hour post infection incubation period based on successful differentiation between infectious and noninfectious viruses and significant and consistent viral replication rates. Ultraviolet disinfection studies were performed to validate the ICC-qRTPCR assay. Using the optimized assay, three-log microbial inactivation was achieved at UV doses of 30–44, 28–42, and 28–29 mJ/cm2 for coxsackievirus B6, echovirus 12, and poliovirus 1, respectively. These results compare favorably to side-by-side assessments using conventional cultural techniques and values previously reported in the literature. This indicates that ICC-qRTPCR is a practical alternative for the simultaneous quantification of enteroviruses in disinfection studies.

Comments

Water Science & Technology, Vol. 61, No. 2 (2010): 375-387. DOI.

Brooke Mayer was affiliated with the Arizona State University at the Tempe Campus at the time of publication.

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