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Development and Evaluation of EPA Method 1615 for Detection of Enterovirus and Norovirus in Water
Citation:
Cashdollar, J., N. Brinkman, S. Griffin, B. Mcminn, E. Rhodes, E. Varughese, A. Grimm, S. Parshionikar, L. Wymer, AND Shay Fout. Development and Evaluation of EPA Method 1615 for Detection of Enterovirus and Norovirus in Water. APPLIED AND ENVIRONMENTAL MICROBIOLOGY. American Society for Microbiology, Washington, DC, 79(1):215-223, (2013).
Impact/Purpose:
The purpose of this study was to describe the developement and evaluation of EPA Method 1615 using groundwater, reagent-grade water, and surface water samples.
Description:
The U.S. EPA developed a sample concentration and preparation assay in conjunction with the Total Culturable Virus Assay for concentrating and measuring culturable viruses in source and drinking waters as part of the Information Collection Rule promulgated in 1996. In an effort to improve upon this method, the U.S. EPA recently developed Method 1615: Measurement of Enterovirus and Norovirus Occurrence in Water by Culture and RT-qPCR. Method 1615 uses a culturable virus assay with reduced equipment and labor costs compared to the ICR virus method and introduces a new molecular assay for detection of enteroviruses and noroviruses by reverse transcription-quantitative polymerase chain reaction. In this study, we describe the optimization of several new components of the molecular assay and examine virus recovery from ground, reagent, and surface waters seeded with poliovirus 3 and murine norovirus. For the culturable virus and molecular assays, respectively, mean poliovirus recoveries using the complete method were 58% and 20% in groundwaters, 122% and 39% using low titer spikes in reagent-grade water, 42% and 48% using high titer spikes in reagent-grade water, and 11% and 10% in surface water with high turbidity. Murine norovirus recoveries by the molecular assay were 29% in groundwaters, less than 8% in both low and high spikes in reagent-grade water, and were 6% in surface water with high turbidity. This study demonstrates the effectiveness of Method 1615 for use in groundwater samples.
