Skip to main content

Advertisement

SpringerLink
Log in

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

  • Original Paper
  • Published:
Food and Environmental Virology Aims and scope Submit manuscript

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.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Similar content being viewed by others

References

  • 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.

    Article  PubMed  CAS  Google Scholar 

  • 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.

    PubMed  CAS  Google Scholar 

  • 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.

    Article  PubMed  CAS  Google Scholar 

  • Choi, W. (2005). Establishment of detection method for norovirus, Technical report (pp. 16–17). Seoul: Korea Food & Drug Administration.

  • 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.

    Article  PubMed  CAS  Google Scholar 

  • Fino, V. R., & Kniel, K. E. (2008). Comparative recovery of foodborne viruses from fresh produce. Foodborne Pathogens and Disease, 5, 819–825.

    Article  PubMed  Google Scholar 

  • 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 

  • Flore, A. E. (2004). Hepatitis A transmitted by food. Clinical Infectious Disease, 38, 705–715.

    Article  Google Scholar 

  • 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.

    Article  PubMed  CAS  Google Scholar 

  • 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 

  • 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.

    CAS  Google Scholar 

  • 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.

    PubMed  CAS  Google Scholar 

  • 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.

    PubMed  CAS  Google Scholar 

  • 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.

    Article  PubMed  CAS  Google Scholar 

  • 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.

    Article  PubMed  CAS  Google Scholar 

  • Koopmans, M., & Duizer, E. (2004). Foodborne viruses: An emerging problem. International Journal of Food Microbiology, 90, 23–41.

    Article  PubMed  Google Scholar 

  • Koopmans, M., von Bonsdorff, C. H., Vinje, J., de Medici, D., & Monroe, S. (2002). Foodborne viruses. FEMS Microbiology Reviews, 26, 187–205.

    PubMed  CAS  Google Scholar 

  • 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.

    Article  PubMed  CAS  Google Scholar 

  • 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.

    PubMed  CAS  Google Scholar 

  • 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.

    CAS  Google Scholar 

  • 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.

    Article  PubMed  CAS  Google Scholar 

  • 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.

    Article  PubMed  CAS  Google Scholar 

  • 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 

  • 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.

    Article  PubMed  CAS  Google Scholar 

  • 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.

    Article  PubMed  Google Scholar 

  • 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.

    PubMed  Google Scholar 

  • 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.

    PubMed  CAS  Google Scholar 

Download references

Acknowledgments

This study was supported by the Basic Science Research Program through the National Research Foundation of Korea (NRF) funded by the Ministry of Education, Science and Technology (2009-0085532).

Author information

Authors and Affiliations

  1. Department of Food and Nutrition, School of Food Science and Technology, Chung-Ang University, 72-1, Nae-ri, Daeduck-myun, Ansung-si, Kyounggi-do, 456-756, South Korea

    Kang Bum Lee & Changsun Choi

  2. Division of Hepatitis Virus, Korea Center for Disease Control and Prevention, Osong Health Technology Administration Complex, 187 Osongsaengmyeong2, Gangoe, Cheongwon, Chungcheongbuk-do, 363-951, South Korea

    Hyeokjin Lee & Doo-Sung Cheon

  3. School of Food Science and Technology, Chung-Ang University, 72-1, Nae-ri, Daeduck-myun, Ansung-si, Kyounggi-do, 456-756, South Korea

    Sang-Do Ha & Changsun Choi

Authors
  1. Kang Bum Lee
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Hyeokjin Lee
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Sang-Do Ha
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Doo-Sung Cheon
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Changsun Choi
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding authors

Correspondence to Doo-Sung Cheon or Changsun Choi.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Lee, K.B., Lee, H., Ha, SD. et al. Comparative Analysis of Viral Concentration Methods for Detecting the HAV Genome Using Real-Time RT-PCR Amplification. Food Environ Virol 4, 68–72 (2012). https://doi.org/10.1007/s12560-012-9077-x

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s12560-012-9077-x

Keywords

Search

Navigation