Publication Abstract

Development of a PCR-based method for the detection of enteroviruses and hepatitis A virus in molluscan shellfish and its application to polluted field samples

Development of a PCR-based method for the detection of enteroviruses and hepatitis A virus in molluscan shellfish and its application to polluted field samples

D.N.Lees, K. Henshilwood and S. Butcher

The use of the polymerase chain reaction (PCR) for detection of low levels of enteric viruses in bivalve shellfish is hindered by the presence of potent amplification inhibitors. A procedure previously developed for removing the majority of these amplification inhibitors is applied to the detection of enteroviruses and hepatitis A virus in naturally polluted field samples. Quantification of PCR inhibition showed that PCR sample tolerance ranged from 2 to 4.7g shellfish for highly polluted samples. These results indicate the need for adequate controls for PCR inhibition, particularly for negative samples. Reverse transcription (RT)-PCR results were compared with conventional enterovirus isolation for a range of naturally contaminated shellfish. All enterovirus isolation positive samples were also positive by enterovirus RT-PCR. At one field site shellfish were positive by enterovirus RT-PCR but negative for virus isolation. All shellfish tested were negative for hepatitis A by RT-PCR. The procedure for removal of PCR amplification inhibitors should be equally applicable to the detection of Norwalk and related Small Round Structured Viruses (SRSVs) in shellfish.

Reference:

D.N.Lees, K. Henshilwood and S. Butcher, 1995. Development of a PCR-based method for the detection of enteroviruses and hepatitis A virus in molluscan shellfish and its application to polluted field samples. Water Science and Technology, 31(5-6): 457-464.

D.N.Lees*, K. Henshilwood* and S. Butcher

Water Science and Technology, 31(5-6): 457-464