Food and Environmental Virology

, Volume 2, Issue 4, pp 218–224 | Cite as

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

  • Vincent R. Hill
  • Bonnie Mull
  • Narayanan Jothikumar
  • Karen Ferdinand
  • Jan Vinjé
Original Paper

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.

Keywords

Norovirus Ground water Ultrafiltration Real-time RT-PCR 

References

  1. 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.CrossRefPubMedGoogle Scholar
  2. 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
  3. 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.CrossRefPubMedGoogle Scholar
  4. Gabrieli, R., Maccari, F., Ruta, A., Paná, A., & Divizia, M. (2009). Norovirus detection in groundwater. Food and Environmental Virology, 1, 92–96.CrossRefGoogle Scholar
  5. Gentry, J., Vinje, J., Guadagnoli, D., & Lipp, E. K. (2009a). Norovirus distribution within an estuarine environment. Applied and Environmental Microbiology, 75, 5474–5480.CrossRefPubMedGoogle Scholar
  6. 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.CrossRefPubMedGoogle Scholar
  7. Glass, R. I., Parashar, U. D., & Estes, M. K. (2009). Norovirus gastroenteritis. New England Journal of Medicine, 361, 1776–1785.CrossRefPubMedGoogle Scholar
  8. Guttman-Bass, N., & Catalano-Sherman, J. (1986). Humic-acid interference with virus recovery by electropositive microporous filters. Applied and Environmental Microbiology, 52, 556–561.PubMedGoogle Scholar
  9. 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.CrossRefPubMedGoogle Scholar
  10. 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.CrossRefPubMedGoogle Scholar
  11. 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.CrossRefPubMedGoogle Scholar
  12. 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.CrossRefPubMedGoogle Scholar
  13. 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.CrossRefGoogle Scholar
  14. 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.CrossRefPubMedGoogle Scholar
  15. 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.CrossRefPubMedGoogle Scholar
  16. 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.CrossRefPubMedGoogle Scholar
  17. 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.CrossRefPubMedGoogle Scholar
  18. 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.CrossRefPubMedGoogle Scholar
  19. 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
  20. 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.CrossRefPubMedGoogle Scholar
  21. 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.CrossRefPubMedGoogle Scholar
  22. 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.CrossRefPubMedGoogle Scholar
  23. 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.CrossRefPubMedGoogle Scholar
  24. 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.CrossRefPubMedGoogle Scholar
  25. 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.CrossRefPubMedGoogle Scholar
  26. 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.CrossRefPubMedGoogle Scholar
  27. 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.PubMedGoogle Scholar
  28. 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.CrossRefPubMedGoogle Scholar
  29. 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

Copyright information

© Springer Science+Business Media, LLC (outside the USA) 2010

Authors and Affiliations

  • Vincent R. Hill
    • 1
  • Bonnie Mull
    • 1
    • 3
  • Narayanan Jothikumar
    • 1
  • Karen Ferdinand
    • 2
    • 3
  • Jan Vinjé
    • 2
  1. 1.Division of Foodborne, Waterborne, and Environmental DiseasesCenters for Disease Control and Prevention, National Center for Emerging and Zoonotic Infectious DiseasesAtlantaUSA
  2. 2.Division of Viral DiseasesCDC, National Center for Immunization and Respiratory DiseasesAtlantaUSA
  3. 3.Atlanta Research and Education FoundationAtlantaUSA

Personalised recommendations