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.
| Original language | English (US) |
|---|---|
| Pages (from-to) | 375-387 |
| Number of pages | 13 |
| Journal | Water Science and Technology |
| Volume | 61 |
| Issue number | 2 |
| DOIs | |
| State | Published - 2010 |
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Keywords
- Enteroviruses
- Integrated cell culture quantitative PCR
- UV disinfection
ASJC Scopus subject areas
- Environmental Engineering
- Water Science and Technology
Cite this
Development and validation of an integrated cell culture-qRTPCR assay for simultaneous quantification of coxsackieviruses, echoviruses, and polioviruses in disinfection studies. / Mayer, B. K.; Ryu, H.; Gerrity, D.; Abbaszadegan, Morteza.
In: Water Science and Technology, Vol. 61, No. 2, 2010, p. 375-387.Research output: Contribution to journal › Article
}
TY - JOUR
T1 - Development and validation of an integrated cell culture-qRTPCR assay for simultaneous quantification of coxsackieviruses, echoviruses, and polioviruses in disinfection studies
AU - Mayer, B. K.
AU - Ryu, H.
AU - Gerrity, D.
AU - Abbaszadegan, Morteza
PY - 2010
Y1 - 2010
N2 - 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.
AB - 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.
KW - Enteroviruses
KW - Integrated cell culture quantitative PCR
KW - UV disinfection
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UR - http://www.scopus.com/inward/citedby.url?scp=77950363676&partnerID=8YFLogxK
U2 - 10.2166/wst.2010.818
DO - 10.2166/wst.2010.818
M3 - Article
VL - 61
SP - 375
EP - 387
JO - Water Science and Technology
JF - Water Science and Technology
SN - 0273-1223
IS - 2
ER -