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DOI10.3791/52646
EPA Method 1615. Measurement of Enterovirus and Norovirus Occurrence in Water by Culture and RT-qPCR. Part III. Virus Detection by RT-qPCR
Fout, G. Shay1; Cashdollar, Jennifer L.1; Griffin, Shannon M.1; Brinkman, Nichole E.1; Varughese, Eunice A.1; Parshionikar, Sandhya U.2
发表日期2016
ISSN1940-087X
期号107
英文摘要

EPA Method 1615 measures enteroviruses and noroviruses present in environmental and drinking waters. This method was developed with the goal of having a standardized method for use in multiple analytical laboratories during monitoring period 3 of the Unregulated Contaminant Monitoring Rule. Herein we present the protocol for extraction of viral ribonucleic acid ( RNA) from water sample concentrates and for quantitatively measuring enterovirus and norovirus concentrations using reverse transcription-quantitative PCR (RT-qPCR). Virus concentrations for the molecular assay are calculated in terms of genomic copies of viral RNA per liter based upon a standard curve. The method uses a number of quality controls to increase data quality and to reduce interlaboratory and intralaboratory variation. The method has been evaluated by examining virus recovery from ground and reagent grade waters seeded with poliovirus type 3 and murine norovirus as a surrogate for human noroviruses. Mean poliovirus recoveries were 20% in groundwaters and 44% in reagent grade water. Mean murine norovirus recoveries with the RT-qPCR assay were 30% in groundwaters and 4% in reagent grade water.


英文关键词Environmental Sciences;Issue 107;virus;waterborne;detection;occurrence;total culturable virus assay;RT-qPCR
语种英语
WOS记录号WOS:000368577400001
来源期刊JOVE-JOURNAL OF VISUALIZED EXPERIMENTS
来源机构美国环保署
文献类型期刊论文
条目标识符http://gcip.llas.ac.cn/handle/2XKMVOVA/62577
作者单位1.US EPA, Natl Exposure Res Lab, Washington, DC 20005 USA;
2.US EPA, Tech Serv Ctr, Off Ground Water & Drinking Water, Washington, DC USA
推荐引用方式
GB/T 7714
Fout, G. Shay,Cashdollar, Jennifer L.,Griffin, Shannon M.,et al. EPA Method 1615. Measurement of Enterovirus and Norovirus Occurrence in Water by Culture and RT-qPCR. Part III. Virus Detection by RT-qPCR[J]. 美国环保署,2016(107).
APA Fout, G. Shay,Cashdollar, Jennifer L.,Griffin, Shannon M.,Brinkman, Nichole E.,Varughese, Eunice A.,&Parshionikar, Sandhya U..(2016).EPA Method 1615. Measurement of Enterovirus and Norovirus Occurrence in Water by Culture and RT-qPCR. Part III. Virus Detection by RT-qPCR.JOVE-JOURNAL OF VISUALIZED EXPERIMENTS(107).
MLA Fout, G. Shay,et al."EPA Method 1615. Measurement of Enterovirus and Norovirus Occurrence in Water by Culture and RT-qPCR. Part III. Virus Detection by RT-qPCR".JOVE-JOURNAL OF VISUALIZED EXPERIMENTS .107(2016).
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