Skip to main content

Advertisement

Springer Nature Link
Log in

Rapid Detection of Human Norovirus in Frozen Raspberries

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

Abstract

Raspberries have lately caused several human norovirus (HuNoV) outbreaks in Europe. In this study, we developed and evaluated for HuNoV reverse transcription (RT)-PCR detection in frozen raspberries extraction methods that have equal sensitivity but are less time-consuming than widely used methods based on polyethylene glycol (PEG) precipitation and chloroform–butanol purification. One method was applied to stored frozen raspberries linked to previous HuNoV outbreaks and berries on sale. In the virus elution-based Method 1, sparkling water eluted viruses most efficiently from the berries. Method 2, based on direct nucleic acid extraction with minor PEG supplement, yielded the highest number of positive findings (4 out of 9) at low virus concentration level of 100 genome copies HuNoV genogroup II per 25 g raspberries. Both methods showed approximately equal sensitivity to a method including PEG precipitation and chloroform–butanol purification. Two naturally contaminated berry samples linked to HuNoV outbreaks in 2006 and 2009 were still positive for HuNoV genogroup I, but all berry products purchased from a local store remained negative for HuNoV. In conclusion, this study presents two efficient and rapid methods which can be used in urgent HuNoV outbreak investigations, since the results of the virus analysis are available in a few hours.

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

Access this article

Log in via an institution

Subscribe and save

Springer+ Basic
$34.99 /Month
  • Get 10 units per month
  • Download Article/Chapter or eBook
  • 1 Unit = 1 Article or 1 Chapter
  • Cancel anytime
Subscribe now

Buy Now

Price includes VAT (Canada)

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2

Similar content being viewed by others

References

  • Anderson, A. D., Garrett, V. D., Sobel, J., Monroe, S. S., Frankhauser, R. L., Schwab, K. J., et al. (2001). Multistate outbreak of norwalk-like virus gastroenteritis associated with a common carter. American Journal of Epidemiology, 154(11), 1013–1019.

    Article  CAS  PubMed  Google Scholar 

  • Baert, L., Mattison, K., Loisy-Hamon, F., Harlow, J., Martyres, A., Lebeau, B., et al. (2011). Review: Norovirus prevalence in Belgian, Canadian and French fresh produce: A threat to human health? International Journal of Food Microbiology, 151, 261–269.

    Article  CAS  PubMed  Google Scholar 

  • 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 

  • Bitler, E., Matthews, J., Dickey, B., Eisenberg, J., & Leon, J. (2013). Norovirus outbreaks: A systematic review of commonly implicated transmission routes and vehicles. Epidemiology and Infection, 141, 1563–1571.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Bouwknegt, M., Verhaelen, K., Rzezutka, A., Kozyra, I., Maunula, L., von Bonsdorff, C.-H., et al. (2015). Quantitative farm-to-fork risk assessment model for norovirus and hepatitis A virus in European leafy green vegetable and berry fruit supply chains. International Journal of Food Microbiology, 198, 50–58.

    Article  PubMed  Google Scholar 

  • Boxman, I., Tilburg, J., te Loeke, N., 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(2), 504–508.

    Article  CAS  PubMed  Google Scholar 

  • Butot, S., Putallaz, T., & Sanchez, G. (2007). Procedure for rapid concentration and detection of enteric viruses from berries and vegetables. Applied and Environmental Microbiology, 73(1), 186–192.

    Article  CAS  PubMed  Google Scholar 

  • Butot, S., Zuber, S., & Baert, L. (2014). Sample preparation prior to molecular amplification: Complexities and opportunities. Current Opinion on Virology, 4, 66–70.

    Article  CAS  Google Scholar 

  • Carter, M. J. (2005). Enterically infecting viruses: Pathogenicity, transmission and significance for food and waterborne infection. Journal of Applied Microbiology, 98, 1354–1380.

    Article  CAS  PubMed  Google Scholar 

  • Cheong, S., Lee, C., Choi, W. C., Lee, C.-H., & Kim, S.-J. (2009). Concentration method for the detection of enteric viruses from large volumes of foods. Journal of Food Protection, 72(9), 2001–2005.

    Article  PubMed  Google Scholar 

  • Cotterelle, B., Drougard, C., Rolland, J., Becamel, M., Boudon, M., Piende, S., et al. (2005). Outbreak of norovirus infection associated with the consumption of frozen raspberries, France, March 2005. Eurosurveillance, 10(17), 2690.

    Google Scholar 

  • Coudray-Meunier, C., Fraisse, A., Martin-Latil, S., Guillier, L., Delannoy, S., Fach, P., et al. (2015). A comparative study of digital RT-PCR and RT-qPCR for quantification of hepatitis A virus and norovirus in lettuce and water samples. International Journal of Food Microbiology, 201, 17–26.

    Article  CAS  PubMed  Google Scholar 

  • de Abreu Corrêa, A., & Miagostovich, M. P. (2013). Optimization of an adsorption-elution method with a negatively charged membrane to recover norovirus from lettuce. Food and Environmental Virology, 5, 144–149.

    Article  Google Scholar 

  • De Keuckelaere, A., Li, D., Deliens, B., Stals, A., & Uyttendaele, M. (2015). Batch testing for noroviruses in frozen raspberries. International Journal of Food Microbiology, 192, 43–50.

    Article  PubMed  Google Scholar 

  • Dolin, R., Blacklow, N., DuPont, H., Buscho, R., Wyatt, R., Kasel, J., et al. (1972). Biological properties of norwalk agent of acute infectious nonbacterial gastroenteritis. Proceedings of the Society for Experimental Biology and Medicine, 140, 578–583.

    Article  CAS  PubMed  Google Scholar 

  • Dubois, E., Agier, C., Traore, O., Hennechart, C., Merle, G., Cruciere, C., et al. (2002). Modified concentration method for the detection of enteric viruses on fruits and vegetables by reverse transcriptase-polymerase chain reaction or cell culture. Journal of Food Protection, 65(12), 1962–1969.

    Article  CAS  PubMed  Google Scholar 

  • Dubois, E., Hennechart, C., Deboosere, N., Merle, G., Legeay, O., Burger, C., et al. (2006). Intra-laboratory validation of a concentration method adapted for the enumeration of infectious F-specific RNA coliphage, enterovirus, and hepatitis A virus from inoculated leaves of salad vegetables. International Journal of Food Microbiology, 108, 164–171.

    Article  CAS  PubMed  Google Scholar 

  • European Commission (2014). RASFF, the rapid alert system for food and feed, 2013 Annual Report.

  • European Commission (2015). RASFF, the rapid Alert system for food and feed, 2014 Annual Report.

  • Falkenhorst, G., Krusell, L., Lisby, M., Madsen, S., Böttinger, B., & Mølbak, K. (2005). Imported frozen raspberries cause a series of norovirus outbreaks in Denmark, 2005. Eurosurveillance, 10(38), 2795.

    Google Scholar 

  • Fraisse, A., Coudray-Meunier, C., Martin-Latil, S., Hennechart-Collette, C., Delannoy, S., Fach, P., et al. (2017). Digital RT-PCR method for hepatitis A virus and norovirus quantification in soft berries. International Journal of Food Microbiology, 243, 36–45.

    Article  CAS  PubMed  Google Scholar 

  • Fraisse, A., Temmam, S., Deboosere, N., Guillier, L., Delobel, A., Maris, P., et al. (2011). Comparison of chlorine and peroxyacetic-based disinfectant to inactivate feline calicivirus, murine norovirus and hepatitis A VIRUS on lettuce. International Journal of Food Microbiology, 151, 98–104.

    Article  CAS  PubMed  Google Scholar 

  • Fumian, T., Leite, J., Marin, V., & Miagostovich, M. (2009). A rapid procedure for detecting noroviruses from cheese and fresh lettuce. Journal of Virological Methods, 155, 39–43.

    Article  CAS  PubMed  Google Scholar 

  • Guévremont, E., Brassard, J., Houde, A., Simard, C., & Trottier, Y.-L. (2006). Development of an extraction and concentration procedure and comparison of RT-PCR primer system for the detection of hepatitis A virus and norovirus GII in green onions. Journal of Virological Methods, 134, 130–135.

    Article  PubMed  Google Scholar 

  • Hewitt, J., Rivera-Aban, M., & Greening, G. (2009). Evaluation of murine norovirus as a surrogate for human norovirus and hepatitis A virus in heat inactivation studies. Journal of Applied Microbiology, 107, 65–71.

    Article  CAS  PubMed  Google Scholar 

  • Hjertqvist, M., Johansson, A., Svensson, N., Åbom, P.-E., Magnusson, C., Olsson, M., et al. (2006). Four outbreaks of norovirus gastroenteritis after consuming raspberries, Sweden, June–August 2006. Eurosurveillance, 11(36), 3038.

    Google Scholar 

  • Jones, T. H., Brassard, J., Johns, M. W., & Gagne, M.-J. (2009). The effect of pre-treatment and sonication of centrifugal ultrafiltration devices on virus recovery. Journal of Virological Methods, 161, 199–204.

    Article  CAS  PubMed  Google Scholar 

  • Kim, H.-Y., Kwak, I.-S., Hwang, I.-G., & Ko, G. P. (2008). Optimization of methods for detecting norovirus on various fruit. Journal of Virological Methods, 153, 104–110.

    Article  CAS  PubMed  Google Scholar 

  • Kokkinos, P., Kozyra, I., Lazic, S., Bouwknegt, M., Rutjes, S., Willem, K., et al. (2012). Harmonised investigation of the occurrence of human enteric viruses in the leafy green vegetable supply chain in three European countries. Food and Environmental Virology, 4, 179–191.

    Article  CAS  PubMed  Google Scholar 

  • Le Guyader, F. S., Mittelholzer, C., Haugarreau, L., Hedlund, K.-O., Alsterlund, R., Pommepuy, M., et al. (2004). Detection of noroviruses in raspberries associated with a gastroenteritis outbreak. International Journal of Food Microbiology, 97, 179–186.

    Article  PubMed  Google Scholar 

  • Lee, H.-M., Kwon, J., Choi, J.-S., Lee, K.-H., Yang, S., Ko, S.-M., et al. (2013). Rapid detection of norovirus from fresh lettuce using immunomagnetic separation and a quantum dots assay. Journal of Food Protection, 76(4), 707–711.

    Article  CAS  PubMed  Google Scholar 

  • Mathijs, E., Stals, A., Baert, L., Botteldoorn, N., Denayer, S., Mauroy, A., et al. (2012). A review of known and hypothetical transmission routes for noroviruses. Food and Environmental Virology, 4, 131–152.

    Article  PubMed  Google Scholar 

  • Matthews, J., Dickey, B., Miller, R., Felzer, J., Dawson, B., Lee, A., et al. (2012). The epidemiology of published norovirus outbreaks: A systematic review of risk factors associated with attack rate and genogroup. Epidemiology and Infection, 140, 1161–1172.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Maunula, L., Kaupke, A., Vasickova, P., Söderberg, K., Kozyra, I., Lazic, S., et al. (2013). Tracking enteric viruses in the European berry fruit supply chain. International Journal of Food Microbiology, 167, 177–185.

    Article  PubMed  Google Scholar 

  • Maunula, L., Roivainen, M., Keränen, M., Mäkelä, S., Söderberg, K., Summa, M., et al. (2009). Detection of human norovirus from frozen raspberries in a cluster of gastroenteritis outbreaks. Eurosurveillance, 14(49), 19435.

    PubMed  Google Scholar 

  • Mokhtari, A., & Jaykus, L.-A. (2009). Quantitative exposure model for the transmission of norovirus in retail food preparation. International Journal of Food Microbiology, 133, 38–47.

    Article  PubMed  Google Scholar 

  • Morillo, S., Luchs, A., Cilli, A., & Timenetsky, M. (2012). Rapid detection of norovirus in naturally contaminated food: Foodborne gastroenteritis outbreak on a Cruise Ship in Brazil, 2010. Food and Environmental Virology, 4, 124–129.

    Article  PubMed  Google Scholar 

  • Park, H., Kim, M., & Ko, G. (2010). Evaluation of various methods for recovering human norovirus and murine norovirus from vegetables and ham. Journal of Food Protection, 73(9), 1651–1657.

    Article  CAS  PubMed  Google Scholar 

  • Perrin, A., Loutreul, J., Boudaud, N., Bertrand, I., & Ganzer, C. (2015). Rapid, simple and efficient method for detection of viral genomes on raspberries. Journal of Virological Methods, 224, 95–101.

    Article  CAS  PubMed  Google Scholar 

  • Rönnqvist, M., Aho, E., Mikkelä, A., Ranta, J., Tuominen, P., Rättö, M., et al. (2014). Norovirus transmission between hands, gloves, utensils, and fresh produce during simulated food handling. Applied and Environmental Microbiology, 80(17), 5403–5410.

    Article  PubMed  PubMed Central  Google Scholar 

  • Rutjes, S. A., Lodder-Verschoor, F., van der Poel, W. H. M., van Duijnhoven, Y. T. H. P., & de Roda, Husman A. M. (2006). Detection of noroviruses in foods: A study on virus extraction procedures in foods implicated in outbreaks of human gastroenteritis. Journal of Food Protection, 69(8), 1949–1956.

    Article  PubMed  Google Scholar 

  • Rzezutka, A., Alotaibi, M., D’Agostino, M., & Cook, N. (2005). A centrifugation-based method for extraction of norovirus from raspberries. Journal of Food Protection, 68(9), 1923–1925.

    Article  CAS  PubMed  Google Scholar 

  • Sarvikivi, E., Roivainen, M., Maunula, L., Niskanen, T., Korhonen, T., Lappalainen, M., et al. (2012). Multiple norovirus outbreaks linked to imported frozen raspberries. Epidemiology and Infection, 140, 260–267.

    Article  CAS  PubMed  Google Scholar 

  • Schwab, K., Neill, F., Fankhauser, R., Daniels, N., Monroe, S., Bergmire-Sweat, D., et al. (2000). Development of methods to detect “norwalk-like viruses” (NLVs) and hepatitis A virus in delicatessen foods: Application to a food-borne NLV outbreak. Applied and Environmental Microbiology, 66(1), 213–218.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Stals, A., Baert, L., Jasson, V., van Coillie, E., & Uyttendaele, M. (2011a). Screening of fruit products for norovirus and the difficulty of interpreting positive PCR results. Journal of Food Protection, 74(3), 425–431.

    Article  PubMed  Google Scholar 

  • Stals, A., Baert, L., van Coillie, E., & Uyttendaele, M. (2011b). Evaluation of a norovirus detection methodology for soft red fruits. Food Microbiology, 28, 52–58.

    Article  PubMed  Google Scholar 

  • Stals, A., Baert, L., van Coillie, E., & Uyttendaele, M. (2012). Extraction of food-borne viruses from food samples: A review. International Journal of Food Microbiology, 153, 1–9.

    Article  PubMed  Google Scholar 

  • Stals, A., Uyttendaele, M., Baert, L., & van Coillie, E. (2013). Norovirus transfer between foods and food contact materials. Journal of Food Protection, 76(7), 1202–1209.

    Article  PubMed  Google Scholar 

  • Summa, M., von Bonsdorff, C.-H., & Maunula, L. (2012a). Evaluation of four virus recovery methods for detecting noroviruses on fresh lettuce, sliced ham, and frozen raspberries. Journal of Virological Methods, 183, 154–160.

    Article  CAS  PubMed  Google Scholar 

  • Summa, M., von Bonsdorff, C.-H., & Maunula, L. (2012b). Pet dogs—A transmission route for human noroviruses? Journal of Clinical Virology, 53, 244–247.

    Article  PubMed  Google Scholar 

  • Teunis, P., Moe, C., Liu, P., Miller, S., Lindesmith, L., Baric, R., et al. (2008). Norwalk virus: How infectious is it? Journal of Medical Virology, 80, 1468–1476.

    Article  PubMed  Google Scholar 

  • Tuladhar, E., Hazeleger, W., Koopmans, M., Zwietering, M., Duizer, E., & Beumer, R. (2013). Transfer of noroviruses between fingers and fomites and food products. International Journal of Food Microbiology, 167, 346–352.

    Article  PubMed  Google Scholar 

  • Verhaelen, K., Bouwknegt, M., Carratala, A., Lodder-Verschoor, F., Diez-Valcarce, M., Rodriguez-Lazaro, D., et al. (2013). Virus transfer proportions between gloved fingertips, soft berries, and lettuce, an associated health risks. International Journal of Food Microbiology, 166, 419–425.

    Article  PubMed  Google Scholar 

  • White, P. (2014). Evolution of norovirus. Clinical Microbiology & Infection, 20, 741–745.

    Article  CAS  Google Scholar 

  • Wilson, I. G. (1997). Inhibition and facilitation of nucleic acid amplification. Applied and Environmental Microbiology, 63(10), 3741–3751.

    CAS  PubMed  PubMed Central  Google Scholar 

  • Zheng, D.-U., Ando, T., Fankhauser, R., Beard, R., Glass, R., & Monroe, S. (2006). Norovirus classification and proposed strain nomenclature. Virology, 346, 312–323.

    Article  CAS  PubMed  Google Scholar 

Download references

Acknowledgements

We thank K. Söderberg for her valuable contribution during the method design and C.H. von Bonsdorff for his specialist advice when proceeding this study.

Funding

The work was funded by the Academy of Finland (Grant Number 1122782), the Walter Ehrström Foundation and the Finnish Foundation of Veterinary Research.

Author information

Authors and Affiliations

  1. Department of Food Hygiene and Environmental Health, Faculty of Veterinary Medicine, University of Helsinki, PO Box 66, 00014, Helsinki, Finland

    Maija Summa & Leena Maunula

Authors
  1. Maija Summa
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Leena Maunula
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Maija Summa.

Ethics declarations

Conflict of interest

The authors declare that they have no conflict of interest.

Rights and permissions

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Summa, M., Maunula, L. Rapid Detection of Human Norovirus in Frozen Raspberries. Food Environ Virol 10, 51–60 (2018). https://doi.org/10.1007/s12560-017-9321-5

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s12560-017-9321-5

Keywords

Search

Navigation