EPA Method 1615 Measurement of Enterovirus and Norovirus Occurrence in Water by Culture and RT-qPCR (January 2012)
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7C65EFB2C8FA4F159FFDCCFC8061947F |
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文件大小(MB)： |
1.06 |
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页数： |
102 |
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日期： |
2012-3-5 |
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文本摘录（文本识别可能有误，但文件阅览显示及打印正常，pdf文件可进行文字搜索定位）：
Version 1.1 Revised January 2012,Cover:,Left picture: Prairie Du Sac, WI Pump house, courtesy of Dr. Mark Borchardt,Middle picture: norovirus, courtesy of Fred P. Williams; Bar = 50 nanometers,Right picture: poliovirus, courtesy of Fred P. Williams; Bar = 50 nanometers,EPA/600/R-10/181,Revised January 2012,Method 1615,Measurement of Enterovirus and Norovirus,Occurrence in Water by Culture and RT-qPCR,Version 1.1,G.S. Fout1, N.E. Brinkman1,*, J.L. Cashdollar1,*, S.M. Griffin1,*, B.R. McMinn1,*, E.R. Rhodes1,*, E.A. Varughese1,*, M.R. Karim1, A.C. Grimm1, S.K. Spencer2, and M.A. Borchardt2,1 U.S. Environmental Protection Agency, Office of Research and Development,National Exposure Research Laboratory, Cincinnati, OH,2 U.S. Department of Agriculture, Agricultural Research Service, Marshfield, WI,* NEB, JLC, SMG, BRM, ERR, and EAV contributed equally to the,evaluation of Method 1615,National Exposure Research Laboratory,Office of Research and Development,U.S. Environmental Protection Agency,Cincinnati, OH 45268,i,Disclaimer,This method has been reviewed by the U.S. Environmental Protection Agency (EPA)’s Office of Research and Development (ORD) and approved for publication. Mention of trade names or commercial products does not constitute endorsement or recommendation for use. The initial intended use of Method 1615 is to support the nationwide monitoring of enteroviruses and noroviruses under the Unregulated Contaminant Monitoring Regulation (UCMR). The method may however, be adopted in the future for other Safe Drinking Water Act and Clean Water Act purposes.,Acknowledgements,The authors thank Drs. Sandhya Parshionikar, Keya Sen, and Carrie Miller from EPA’s Office of Water and Drs. George Di Giovanni from the Texas AgriLife Research Center, El Paso, TX, Paul Hazelton from the University of Manitoba, Winnipeg, Manitoba, Timothy Straub from the Pacific Northwest National Laboratory, Richland, WA, and Susan Boutros from Analytical Associates, Ithaca, NY for reviewing this manuscript and for providing helpful comments, and Justicia Rhodus from Dynamac Corporation for providing technical edits. Mohammad R. Karim was supported through an appointment to the Research Participation Program at ORD’s National Exposure Research Laboratory, which was administered by the Oak Ridge Institute for Science and Education through an Interagency Agreement between the U.S. Department of Energy and EPA. The authors thank Dr. Skip Virgin of Washington University School of Medicine, St. Louis, MO for murine norovirus.,Questions regarding this method or its application should be addressed to:,G. Shay Fout, Ph.D.,U.S. Environmental Protection Agency,Office of Research and Development,National Exposure Research Laboratory,Microbiological and Chemical Exposure,Assessment Research Division,Cincinnati, OH 45268-1320,(513) 569-7387,(513) 569-7117 (fax),fout.shay@epa.gov,ii,Table of Contents,ABBREVIATIONS v,1.0 SCOPE AND APPLICATION 1,1.1 Background 1,1.2 Method Constraints 3,2.0 SUMMARY OF METHOD .. 3,3.0 DEFINITIONS . 3,4.0 INTERFERENCES 6,4.1 Reagents .. 6,4.2 Matrix Interference 6,4.3 Other Interference .. 6,5.0 SAFETY . 7,5.1 Safety Plan . 7,5.2 Shipment of Field Samples 7,5.3 Chemical Safety .. 7,6.0 EQUIPMENT AND SUPPLIES . 7,6.1 Sample Filtration Apparatus . 8,6.2 Other Equipment and Supplies for Sample Collection, Preservation, and Storage Procedure . 10,6.3 Equipment and Supplies for Quality Assurance Measures . 11,6.4 Equipment and Supplies for the Elution and Organic Flocculation Procedures .. 12,6.5 Equipment and Supplies for the Total Culturable Virus Assay 14,6.6 Equipment and Supplies for the Enterovirus and Norovirus Molecular Assays . 14,6.7 Equipment and Supplies for Sterilization Techniques .. 15,7.0 REAGENTS, MEDIA, AND STANDARDS 16,7.1 Reagents for the Sample Collection, Preservation and Storage Procedure 16,7.2 Reagents for Quality Assurance Measures . 16,7.3 Reagents for the Elution and Organic Flocculation Procedures .. 17,7.4 Reagents for the Total Culturable Virus Assay 19,7.5 Reagents for the Enterovirus and Norovirus Molecular Assays .. 21,7.6 Reagents for Sterilization Techniques .. 22,8.0 QUALITY ASSURANCE .. 22,8.1 Quality Assurance Plan . 22,8.2 Laboratory Personnel . 24,8.3 Laboratory Performance .. 25,8.4 QC Sample Set .. 26,8.5 PT and PE Samples . 27,8.6 Matrix Spike 28,iii,8.7 Record Maintenance .. 29,9.0 SAMPLE COLLECTION, PRESERVATION, AND STORAGE 29,9.1 Field Sample Collection 29,9.2 Shipment of Field Samples . 35,9.3 Laboratory Holding Time and Temperature .. 35,10.0 FILTER ELUTION PROCEDURE 36,10.1 Elution Equipment Setup . 36,10.2 Elution 36,11.0 OR……
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