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Detection of GI and GII Noroviruses in Ground Water Using Ultrafiltration and TaqMan Real-time RT-PCR

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Abstract

Noroviruses (NoVs) are a leading cause of epidemic and sporadic acute gastrointestinal illness globally. These viruses can potentially contaminate rural private wells and non-community drinking water systems, and cause waterborne disease outbreaks related to consumption of contaminated ground water. Detection of NoVs in water samples can be challenging because they are genetically and antigenically diverse, and noncultivable. In the present study, the detection limits of a novel broadly reactive GI assay and an existing GII NoV real-time TaqMan reverse transcriptase-polymerase chain reaction (RT-qPCR) assay in ground water concentrates was determined. Ground water samples (50 l) from two sources (Lawrenceville, GA and Gainesville, FL, USA) were seeded with electron microscopy-enumerated and RT-qPCR quantified NoV and concentrated using hollow-fiber ultrafiltration (UF) followed by either polyethylene glycol (PEG) precipitation or microconcentrators. Detection limits for GI NoV ranged from 1 × 104 (GA source) to 2 × 105 (FL source) virus particles in 50 l water samples (corresponding to 200–3,000 particles/l) and 5 × 104 (GA source) to 5 × 105 (FL source) virus particles (corresponding to 1,000–10,000 particles/l) for GII NoV. The reported UF method, sample processing procedures, and RT-qPCR assays should be effective tools for sensitive detection of NoVs in large-volume water samples.

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References

  • Baert, L., Uyttendaele, M., & Debevere, J. (2008). Evaluation of viral extraction methods on a broad range of ready-to-eat foods with conventional and real-time RT-PCR for Norovirus GII detection. International Journal of Food Microbiology, 123, 101–108.

    Article  CAS  PubMed  Google Scholar 

  • Cannon, J., & Vinje, J. (2008). Foodborne viruses. In E. Palombo & C. Kirkwood (Eds.), Viruses in the environment (1st ed., pp. 45–76). Ames: Iowa State Press.

    Google Scholar 

  • Costantini, V., Grenz, L., Fritzinger, A., Lewis, D., Biggs, C., Hale, A., et al. (2010). Diagnostic accuracy and analytical sensitivity of IDEIA norovirus assay for routine screening of human norovirus. Journal of Clinical Microbiology, 48, 2770–2778.

    Article  CAS  PubMed  Google Scholar 

  • Gabrieli, R., Maccari, F., Ruta, A., Paná, A., & Divizia, M. (2009). Norovirus detection in groundwater. Food and Environmental Virology, 1, 92–96.

    Article  Google Scholar 

  • Gentry, J., Vinje, J., Guadagnoli, D., & Lipp, E. K. (2009a). Norovirus distribution within an estuarine environment. Applied and Environmental Microbiology, 75, 5474–5480.

    Article  CAS  PubMed  Google Scholar 

  • Gentry, J., Vinje, J., & Lipp, E. K. (2009b). A rapid and efficient method for quantitation of genogroups I and II norovirus from oysters and application in other complex environmental samples. Journal of Virological Methods, 156, 59–65.

    Article  CAS  PubMed  Google Scholar 

  • Glass, R. I., Parashar, U. D., & Estes, M. K. (2009). Norovirus gastroenteritis. New England Journal of Medicine, 361, 1776–1785.

    Article  CAS  PubMed  Google Scholar 

  • Guttman-Bass, N., & Catalano-Sherman, J. (1986). Humic-acid interference with virus recovery by electropositive microporous filters. Applied and Environmental Microbiology, 52, 556–561.

    CAS  PubMed  Google Scholar 

  • Hamza, I. A., Jurzik, L., Stang, A., Sure, K., Uberla, K., & Wilhelm, M. (2009). Detection of human viruses in rivers of a densly-populated area in Germany using a virus adsorption elution method optimized for PCR analyses. Water Research, 43, 2657–2668.

    Article  CAS  PubMed  Google Scholar 

  • Hernandez-Morga, J., Leon-Felix, J., Peraza-Garay, F., Gil-Salas, B. G., & Chaidez, C. (2009). Detection and characterization of hepatitis A virus and norovirus in estuarine water samples using ultrafiltration—RT-PCR integrated methods. Journal of Applied Microbiology, 106, 1579–1590.

    Article  CAS  PubMed  Google Scholar 

  • Hewitt, J., Bell, D., Simmons, G. C., Rivera-Aban, M., Wolf, S., & Greening, G. E. (2007). Gastroenteritis outbreak caused by waterborne norovirus at a New Zealand ski resort. Applied and Environmental Microbiology, 73, 7853–7857.

    Article  CAS  PubMed  Google Scholar 

  • Hill, V. R., Kahler, A. M., Jothikumar, N., Johnson, T. B., Hahn, D., & Cromeans, T. L. (2007). Multistate evaluation of an ultrafiltration-based procedure for simultaneous recovery of enteric microbes in 100-liter tap water samples. Applied and Environmental Microbiology, 73, 4218–4225.

    Article  CAS  PubMed  Google Scholar 

  • Huffman, D. E., Nelson, K. L., & Rose, J. B. (2003). Calicivirus—an emerging contaminant in water: state of the art. Environmental Engineering Science, 20, 503–515.

    Article  CAS  Google Scholar 

  • Hunt, R. J., Borchardt, M. A., Richards, K. D., & Spencer, S. K. (2010). Assessment of sewer source contamination of drinking water wells using tracers and human enteric viruses. Environmental Science and Technology, 44, 7956–7963.

    Article  CAS  PubMed  Google Scholar 

  • Jothikumar, N., Lowther, J. A., Henshilwood, K., Lees, D. N., Hill, V. R., & Vinje, J. (2005). Rapid and sensitive detection of noroviruses by using TaqMan-based one-step reverse transcription-PCR assays and application to naturally contaminated shellfish samples. Applied and Environmental Microbiology, 71, 1870–1875.

    Article  CAS  PubMed  Google Scholar 

  • Kageyama, T., Kojima, S., Shinohara, M., Uchida, K., Fukushi, S., Hoshino, F. B., et al. (2003). Broadly reactive and highly sensitive assay for Norwalk-like viruses based on real-time quantitative reverse transcription-PCR. Journal of Clinical Microbiology, 41, 1548–1557.

    Article  CAS  PubMed  Google Scholar 

  • Kaplan, J. E., Feldman, R., Campbell, D. S., Lookabaugh, C., & Gary, G. W. (1982). The frequency of a Norwalk-like pattern of illness in outbreaks of acute gastroenteritis. American Journal of Public Health, 72, 1329–1332.

    Article  CAS  PubMed  Google Scholar 

  • Lambertini, E., Spencer, S. K., Bertz, P. D., Loge, F. J., Kieke, B. A., & Borchardt, M. A. (2008). Concentration of enteroviruses, adenoviruses, and noroviruses from drinking water by use of glass wool filters. Applied and Environmental Microbiology, 74, 2990–2996.

    Article  CAS  PubMed  Google Scholar 

  • Liang, J. L., Dziuban, E. J., Craun, G. F., Hill, V., Moore, M. R., Gelting, R. J., et al. (2006). Surveillance for waterborne disease and outbreaks associated with drinking water and water not intended for drinking—United States, 2003–2004. Morbidity and Mortality Weekly Report, 55, 31–65.

    Google Scholar 

  • Lukasik, J., Scott, T. M., Andryshak, D., & Farrah, S. R. (2000). Influence of salts on virus adsorption to microporous filters. Applied and Environmental Microbiology, 66, 2914–2920.

    Article  CAS  PubMed  Google Scholar 

  • Ngazoa, E. S., Fliss, I., & Jean, J. (2008). Quantitative study of persistence of human norovirus genome in water using TaqMan real-time RT-PCR. Journal of Applied Microbiology, 104, 707–715.

    Article  CAS  PubMed  Google Scholar 

  • O’Reilly, C. E., Bowen, A. B., Perez, N. E., Sarisky, J. P., Miller, M. D., Hubbard, B. C., et al. (2007). A waterborne outbreak of gastroenteritis with multiple etiologies among resort island visitors and residents: Ohio, 2004. Clinical Infectious Diseases, 44, 506–512.

    Article  PubMed  Google Scholar 

  • Park, Y., Cho, Y. H., Jee, Y., & Ko, G. (2008). Immunomagnetic separation combined with real-time reverse transcriptase PCR assays for detection of norovirus in contaminated food. Applied and Environmental Microbiology, 74, 4226–4230.

    Article  CAS  PubMed  Google Scholar 

  • Polaczyk, A. L., Jothikumar, N., Cromeans, T. L., Hahn, D., Roberts, J. M., Amburgey, J. E., et al. (2008). Ultrafiltration-based techniques for rapid and simultaneous concentration of multiple microbe classes from 100-L tap water samples. Journal of Microbiological Methods, 73, 92–99.

    Article  CAS  PubMed  Google Scholar 

  • Rolfe, K. J., Parmar, S., Mururi, D., Wreghitt, T. G., Jalal, H., Zhang, H., et al. (2007). An internally controlled, one-step, real-time RT-PCR assay for norovirus detection and genogrouping. Journal of Clinical Virology, 39, 318–321.

    Article  CAS  PubMed  Google Scholar 

  • Rutjes, S. A., van den Berg, H., Lodder, W. J., & Husman, A. M. D. (2006). Real-time detection of noroviruses in surface water by use of a broadly reactive nucleic acid sequence-based amplification assay. Applied and Environmental Microbiology, 72, 5349–5358.

    Article  CAS  PubMed  Google Scholar 

  • Sobsey, M. D., & Glass, J. S. (1984). Influence of water quality on enteric virus concentration by microporous filter methods. Applied and Environmental Microbiology, 47, 956–960.

    CAS  PubMed  Google Scholar 

  • Wolf, S., Williamson, W. M., Hewitt, J., Rivera-Aban, M., Lin, S., Ball, A., et al. (2007). Sensitive multiplex real-time reverse transcription-PCR assay for the detection of human and animal noroviruses in clinical and environmental samples. Applied and Environmental Microbiology, 73, 5464–5470.

    Article  CAS  PubMed  Google Scholar 

  • Yoder, J., Roberts, V., Craun, G. F., Hill, V., Hicks, L. A., Alexander, N. T., et al. (2008). Surveillance for waterborne disease and outbreaks associated with drinking water and water not intended for drinking—United States, 2005–2006. MMWR Surveillance Summaries, 57, 39–62.

    Google Scholar 

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Acknowledgments

We thank Charles Humphrey (CDC) for performing electron microscopy. This publication was supported in part by funds made available through CDCs Coordinating Office for Terrorism Preparedness and Emergency Response. The use of trade names and names of commercial sources is for identification only and does not imply endorsement by the CDC or the U.S. Department of Health and Human Services. The findings and conclusions in this presentation are those of the authors and do not necessarily represent those of the CDC. This article received clearance through the appropriate channels at the CDC prior to submission.

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Authors and Affiliations

  1. Division of Foodborne, Waterborne, and Environmental Diseases, Centers for Disease Control and Prevention, National Center for Emerging and Zoonotic Infectious Diseases, 1600 Clifton Road NE, Mail Stop D-66, Atlanta, GA, 30329, USA

    Vincent R. Hill, Bonnie Mull & Narayanan Jothikumar

  2. Division of Viral Diseases, CDC, National Center for Immunization and Respiratory Diseases, Atlanta, GA, USA

    Karen Ferdinand & Jan Vinjé

  3. Atlanta Research and Education Foundation, Atlanta, GA, USA

    Bonnie Mull & Karen Ferdinand

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  1. Vincent R. Hill
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  2. Bonnie Mull
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  3. Narayanan Jothikumar
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  4. Karen Ferdinand
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  5. Jan Vinjé
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Correspondence to Vincent R. Hill.

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Hill, V.R., Mull, B., Jothikumar, N. et al. Detection of GI and GII Noroviruses in Ground Water Using Ultrafiltration and TaqMan Real-time RT-PCR. Food Environ Virol 2, 218–224 (2010). https://doi.org/10.1007/s12560-010-9049-y

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  • DOI: https://doi.org/10.1007/s12560-010-9049-y

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