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DOI | 10.3791/52646 |
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 |
ISSN | 1940-087X |
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
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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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