Advertisement

Detection of Enteric Viruses in Shellfish

  • Gary P. Richards
  • Gail E. Greening
Chapter

Abstract

Norovirus and hepatitis A virus contamination are significant threats to the safety of shellfish and other foods. Methods for the extraction and assay of these viruses from shellfish are complex, time consuming, and technically challenging. Here, we itemize some of the salient points in extracting viruses from shellfish tissues and analyzing the extracts. A more comprehensive treatment of these topics will soon be available in: Richards GP, Cliver DO, Greening GE (in press) Foodborne viruses. Compendium of methods for the microbiological examination of foods, 5th edn. American Public Health Association, Washington, DC. Here, we describe some of the basic principles of virus extraction and molecular assay of viruses in shellfish, the importance of controls, and the limitations of both the extraction and assay methods. Recent efforts to validate methods for virus detection in shellfish center on a method currently in use in Canada and another method under validation in the European Union. The availability of such methods is important in tracking sources of outbreaks and may lead to the establishment of routine monitoring programs.

Keywords

Enteric Virus Internal Amplification Control Canadian Food Inspection Agency Shellfish Tissue Guanidinium Isothiocyanate 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.

References

  1. Anon (2010) CEN/TC275/TAG4 draft standards 2010: microbiology of food and animal feeding stuffs – Horizontal method for the detection of hepatitis A virus and norovirus in food using real-time RT-PCR – Part 1: method for quantitative determination, N501. 2010-05-07, version 8, Brussels, BelgiumGoogle Scholar
  2. Greening GE, Hewitt J (2008) Norovirus detection in shellfish using a rapid, sensitive virus recovery method and real-time RT-PCR detection protocol. Food Anal Method 1:109–118CrossRefGoogle Scholar
  3. Harrington PR, Vinje J, Moe CL, Baric RS (2004) Norovirus capture with histo-blood group antigens reveals novel virus-ligand interactions. J Virol 78:3035–3045CrossRefGoogle Scholar
  4. Kingsley DH, Richards GP (2001) Rapid and efficient extraction method for reverse transcription-PCR detection of hepatitis A and Norwalk-like viruses in shellfish. Appl Environ Microbiol 67:4152–4157CrossRefGoogle Scholar
  5. Kingsley DH, Meade GK, Richards GP (2002) Detection of both hepatitis A virus and Norwalk-like virus in imported clams associated with food-borne illness. Appl Environ Microbiol 68:3914–3918CrossRefGoogle Scholar
  6. Lees D, CEN-WG6-TAG4 (2010) International standardisation of a method for detection of human pathogenic viruses in molluscan shellfish. Food Environ Virol 2:146–155CrossRefGoogle Scholar
  7. López-Sabater EI, Deng MY, Cliver DO (1997) Magnetic immunoseparation PCR assay (MIPA) for detection of hepatitis A virus (HAV) in American oyster (Crassostrea virginica). Lett Appl Microbiol 24:101–104CrossRefGoogle Scholar
  8. Richards GP (1999) Limitations of molecular biological techniques for assessing the virological safety of foods. J Food Prot 62:691–697Google Scholar
  9. Richards GP, Cliver DO, Greening GE (in press) Foodborne viruses. Compendium of methods for the microbiological examination of foods, 5th edn. American Public Health Association, Washington, DCGoogle Scholar
  10. Tian P, Engelbrektson A, Mandrell R (2008) Two-log increase in sensitivity for detection of norovirus in complex samples by concentration with porcine gastric mucin conjugated to magnetic beads. Appl Environ Microbiol 74:4271–4276CrossRefGoogle Scholar
  11. Trottier Y-L, Houde A, Buenaventura E et al (2010) Concentration of norovirus genogroups I and II from contaminated oysters and their detection using the reverse-transcriptase polymerase chain reaction. Available online at: http://www.hc-sc.gc.ca/fn-an/res-rech/analy-meth/microbio/volume5/opflp_01-eng.php. Accessed 16 Mar 2011

Copyright information

© Springer Science+Business Media Dordrecht 2014

Authors and Affiliations

  1. 1.United States Department of Agriculture, Agricultural Research Service, James W.W. Baker CenterDelaware State UniversityDoverUSA
  2. 2.Food Group, Kenepuru Science CentreInstitute of Environmental Science and Research Ltd. (ESR)PoriruaNew Zealand

Personalised recommendations