Development and validation of an integrated cell culture-qRTPCR assay for simultaneous quantification of coxsackieviruses, echoviruses, and polioviruses in disinfection studies
Document Type
Article
Publication Date
2010
Publication Title
Water Science and Technology
Volume
61
Issue
2
First page number:
375
Last page number:
387
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/cm(2) 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.
Keywords
Coxsackieviruses; Disinfection and disinfectants; ECHO viruses; Enteroviruses; Poliovirus
Disciplines
Environmental Engineering | Environmental Public Health | Environmental Sciences | Other Engineering | Public Health | Virus Diseases
Language
English
Permissions
Use Find in Your Library, contact the author, or use interlibrary loan to garner a copy of the article. Publisher copyright policy allows author to archive post-print (author’s final manuscript). When post-print is available or publisher policy changes, the article will be deposited
Repository Citation
Mayer, B. K.,
Ryu, H.,
Gerrity, D.,
Abbaszadegan, M.
(2010).
Development and validation of an integrated cell culture-qRTPCR assay for simultaneous quantification of coxsackieviruses, echoviruses, and polioviruses in disinfection studies.
Water Science and Technology, 61(2),
375-387.
http://dx.doi.org/10.2166/wst.2010.819
