Food and Environmental Virology

, Volume 4, Issue 2, pp 68–72

Comparative Analysis of Viral Concentration Methods for Detecting the HAV Genome Using Real-Time RT-PCR Amplification

Original Paper

Abstract

Hepatitis A is a major infectious disease epidemiologically associated with foodborne and waterborne outbreaks. Molecular detection using real-time RT-PCR to detect the hepatitis A virus (HAV) in contaminated vegetables can be hindered by low-virus recoveries during the concentration process and by natural PCR inhibitors in vegetables. This study evaluated three virus concentration methods from vegetables: polyethylene glycol (PEG) precipitation, ultrafiltration (UF), and immunomagnetic separation (IMS). UF was the most efficient concentration method, while PEG and IMS were very low for the recovery rate of HAV. These results demonstrate that UF is the most appropriate method for recovering HAV from contaminated vegetables and that this method combined with the real-time RT-PCR assay may be suitable for routine laboratory use.

Keywords

Hepatitis A virus (HAV) Concentration Detection Ultrafiltration (UF) Immunomagnetic separation (IMS) 

References

  1. Bidawid, S., Farber, J. M., & Sattar, S. A. (2000). Rapid concentration and detection of hepatitis A virus from lettuce and strawberries. Journal of Virological Methods, 88, 175–185.PubMedCrossRefGoogle Scholar
  2. Boxman, I. L., Tilburg, J. J., te Loeke, N. A., Vennema, H., de Boer, E., & Koopmans, M. (2007). An efficient and rapid method for recovery of norovirus from food associated with outbreaks of gastroenteritis. Journal of Food Protection, 70, 504–508.PubMedGoogle Scholar
  3. Butot, S., Putallaz, T., & Sanchez, G. (2007). Procedure for rapid concentration and detection of enteric virus from berries and vegetables. Applied and Environmental Microbiology, 73, 186–192.PubMedCrossRefGoogle Scholar
  4. Choi, W. (2005). Establishment of detection method for norovirus, Technical report (pp. 16–17). Seoul: Korea Food & Drug Administration.Google Scholar
  5. Dubois, E., Hennechart, C., Merle, G., Burger, C., Hmila, N., Ruelle, S., et al. (2007). Detection and quantification by real-time RT-PCR of hepatitis A virus from inoculated tap waters, salad vegetables, and soft fruits: characterization of the method performances. International Journal of Food Microbiology, 117, 141–149.PubMedCrossRefGoogle Scholar
  6. Fino, V. R., & Kniel, K. E. (2008). Comparative recovery of foodborne viruses from fresh produce. Foodborne Pathogens and Disease, 5, 819–825.PubMedCrossRefGoogle Scholar
  7. Fiore, A. E., Wasley, A., & Bell, B. P. (2006). Prevention of hepatitis A through active or passive immunization: recommendations of the Advisory Committee on Immunization Practices (ACIP). Morbidity and Mortality Weekly Report, 55, 1–23.Google Scholar
  8. Flore, A. E. (2004). Hepatitis A transmitted by food. Clinical Infectious Disease, 38, 705–715.CrossRefGoogle Scholar
  9. Gilpatrick, S. G., Schwab, K. J., Estes, M. K., & Atmar, R. L. (2000). Development of an immunomagnetic capture reverse transcription-PCR assay for the detection of Norwalk virus. Journal of Virological Methods, 90, 69–78.PubMedCrossRefGoogle Scholar
  10. Guillois-Becel, Y., Couturier, E., Le Saux, J. C., Roque-Afonso, A. M., Le Guyader, F. S., Le Goas, A., et al. (2009). An oyster-associated hepatitis A outbreak in France in 2007. Euro Surveillance, 14, 1–6.Google Scholar
  11. Ha, S., Woo, G., Hwang, I., & Choi, W. (2009). Development of a virus elution and concentration procedure for detecting norovirus in oysters. Food Science and Biotechnology, 18, 1150–1154.Google Scholar
  12. Hooper, R. R., Juels, C. W., Routenberg, J. A., Harrison, W. O., Kilpatrick, M. E., Kendra, S. J., et al. (1977). An outbreak of type A hepatitis at the Naval Training Center, San Diego: epidemiologic evaluation. American Journal of Epidemiology, 105, 148–155.PubMedGoogle Scholar
  13. Jothikumar, N., Cliver, D. O., & Mariam, T. W. (1998). Immunomagnetic capture PCR for concentration and detection of hepatitis A virus from environmental samples. Applied and Environmental Microbiology, 64, 504–508.PubMedGoogle Scholar
  14. Jothikumar, N., Cromeans, T. L., Sobsey, M. D., & Robertson, B. H. (2005). Development and evaluation of a broadly reactive TaqMan assay for rapid detection of hepatitis A virus. Applied and Environmental Microbiology, 71, 3359–3363.PubMedCrossRefGoogle Scholar
  15. Kittigul, L., Pombubpa, K., Rattanatham, T., Diraphat, P., Utrarachkij, F., Pungchitton, S., et al. (2008). Development of a method for concentrating and detecting rotavirus in oysters. International Journal of Food Microbiology, 122, 204–210.PubMedCrossRefGoogle Scholar
  16. Koopmans, M., & Duizer, E. (2004). Foodborne viruses: An emerging problem. International Journal of Food Microbiology, 90, 23–41.PubMedCrossRefGoogle Scholar
  17. Koopmans, M., von Bonsdorff, C. H., Vinje, J., de Medici, D., & Monroe, S. (2002). Foodborne viruses. FEMS Microbiology Reviews, 26, 187–205.PubMedGoogle Scholar
  18. Love, D. C., Casteel, M. J., Meschke, J. S., & Sobsey, M. D. (2008). Methods for recovery of hepatitis A virus (HAV) and other viruses from processed foods and detection of HAV by nested RT-PCR and TaqMan RT-PCR. International Journal of Food Microbiology, 126, 221–226.PubMedCrossRefGoogle Scholar
  19. McCaustland, K. A., Bond, W. W., Bradley, D. W., Ebert, J. W., & Maynard, J. E. (1982). Survival of hepatitis A virus in feces after drying and storage for one month. Journal of Clinical Microbiology, 16, 957–958.PubMedGoogle Scholar
  20. Moon, A., Hwang, I., & Choi, W. (2009). Development of a virus elution and concentration procedure for detecting norovirus in cabbage and lettuce. Food Science and Biotechnology, 18, 407–412.Google Scholar
  21. Myrmel, M., Rimstad, E., & Wasteson, Y. (2000). Immunomagnetic separation of a Norwalk-like virus (genogroup I) in artificially contaminated environmental water samples. International Journal of Food Microbiology, 62, 17–26.PubMedCrossRefGoogle Scholar
  22. Papafragkou, E., Plante, M., Mattison, K., Bidawid, S., Krthikeyan, K., Farber, J. M., et al. (2008). Rapid and sensitive detection of hepatitis A virus in representative food matrices. Journal of Virological Methods, 147, 177–187.PubMedCrossRefGoogle Scholar
  23. Petriqnani, M., Verhoef, L., van Hunen, R., Swaan, C., van Steenbergen, J., Boxman, I., et al. (2010). A possible foodborne outbreak of hepatitis A in the Netherlands, January–February 2010. Euro Surveillance, 15, pii19512.Google Scholar
  24. Pinto, R. M., Costafreda, M. I., & Bosch, A. (2009). Risk assessment in shelfish-borne outbreaks of hepatitis A. Applied and Environmental Microbiology, 75, 7350–7355.PubMedCrossRefGoogle Scholar
  25. Robesyn, E., De Schrijver, K., Wollants, E., Top, G., Verbeeck, J., & Van Ranst, M. (2009). An outbreak of hepatitis A associated with the consumption of raw beef. Journal of Clinical Virology, 44, 207–210.PubMedCrossRefGoogle Scholar
  26. Rowe, S. L., Tanner, K., & Gregory, J. E. (2009). Hepatitis A outbreak epidemiologically linked to a food handler in Melbourne, Victoria. Communicable Diseases Intelligence, 33, 46–48.PubMedGoogle Scholar
  27. Rzezutka, A., D’Agostino, M., & Cook, N. (2006). An ultracentrifugation-based approach to the detection of hepatitis A virus in soft fruits. International Journal of Food Microbiology, 108, 315–320.PubMedGoogle Scholar

Copyright information

© Springer Science + Business Media, LLC 2012

Authors and Affiliations

  1. 1.Department of Food and Nutrition, School of Food Science and TechnologyChung-Ang UniversityAnsung-siSouth Korea
  2. 2.Division of Hepatitis VirusKorea Center for Disease Control and PreventionCheongwonSouth Korea
  3. 3.School of Food Science and TechnologyChung-Ang UniversityAnsung-siSouth Korea

Personalised recommendations