Use of genomic probes to detect hepatitis A virus and enterovirus RNAs in wild shellfish and relationship of viral contamination to bacterial contamination

Abstract

Genomic probes were used to investigate hepatitis A virus (HAV) and enterovirus RNAs in two types of shellfish from natural beds (Atlantic coast, France). After elution concentration, nucleic acid extracted by proteinase K and purified by phenol-chloroform and ethanol precipitation was assayed by dot blot hybridization. The probes used were a specific HAV probe corresponding to the 3' end (3D polymerase coding region) and an enterovirus probe corresponding to the 5' noncoding region. The method was first tested under experimental conditions by using virus-spiked shellfish before being applied under field conditions. Our results show that shellfish were highly contaminated: enterovirus and HAV RNAs were found in 63 and 67%, respectively, of samples examined with the riboprobes. On the same site, viral (HAV and enterovirus) RNAs were found in a larger fraction of cockles than mussels. Statistical tests of dependence showed no relationship between viral contamination and bacterial contamination (evaluated by fecal coliform counts).

Detection of Norwalk virus and hepatitis A virus in shellfish tissues with the PCR

“…A number of previous reports have described molecular methods for the detection of enteric viruses in shellfish (1, 5-8, 10, 17-19, 27). The use of labeled, virus-specific probes in hybridization assays lacked sufficient sensitivity to detect viruses present in low concentrations in shellfish, although virus could be detected in more heavily contaminated environmental samples (18,27). RT-PCR assays have the potential to overcome the limitations of probe hybridization assays, increasing the theoretical level of detection to as few as one copy of a viral genome.…”

Bacteriophages as indicators of enteric viruses and public health risk in groundwaters

“…It has been suggested that pan-enterovirus probes, general indicators of enteric viral pathogens, should be used in direct tests for HAV, caliciviruses, rotaviruses and astroviruses (Deleon et al 1990;Margolin et al 1991;Schwab et al 1991Schwab et al , 1993. However, there has been little consistent correlation between enteroviruses and HAV occurrence (Dubrou et al 1991;Shieh et al 1991;Leguyader et al 1993Leguyader et al , 1994Tsai et al 1993), and other groups of enteric viruses may show different survival and disinfection-resistant behaviour (Block 1983;Hoff and Akin 1983;Schwartzbrod 1991;West 1991). Therefore, the value of enteroviral detection in predicting a potential public health hazard of viral disease is no better, and may be worse, than that of a faecal bacterial indicator such as E. coli or Enterococci (Metcalf et al 1995).…”

REVERSE TRANSCRIPTASE–POLYMERASE CHAIN REACTION DETECTION OF HEPATITIS A VIRUS IN NATURALLY CONTAMINATED MUSSELS (MYTILUS EDULIS)

“… Sample i (1 < i < 7): HAV‐contaminated mussel samples chosen from site four (Le Guyader et al. 1993).…”

“…To evaluate the efficiency of four enteric viruses, extraction methods (using borate buffer, glycine solution, saline beef and saline beef–PVP) were performed on each of seven highly positive HAV naturally contaminated mussel samples ( M. edulis ) as confirmed by Le Guyader et al. (1993).…”