Detection of hepatitis A virus, rotavirus, and enterovirus in naturally contaminated shellfish and sediment by reverse transcription-seminested PCR.
A reverse transcription-PCR method was developed to detect enterovirus (EV), hepatitis A virus (HAV), and rotavirus (RV) RNAs in shellfish and sediment. The method was first tested under experimental conditions by using virus-spiked shellfish to evaluate assay sensitivity. The use of CC41 cellulose was found to be efficient for removing inhibitors of RV detection. For sediment samples, a Sephadex column was used to allow the detection of EV and HAV RNAs. The specificity of amplified products was controlled by hybridization with digoxigenin-labeled oligoprobes. The method was then applied to naturally contaminated shellfish and sediments. EV, HAV, and RV RNAs were detected in 22, 14, and 20% of the shellfish samples, respectively. No relationship between viral contamination and bacterial contamination was found. When viral RNAs (HAV or EV) were detected in sediments, they were also detected in shellfish.
Le Guyader
F
F
Laboratoire de Microbiologie, Institut Français de Recherche pour l'Exploitation de la Mer, Nantes, France.
Dubois
E
E
Menard
D
D
Pommepuy
M
M
D000818:Animals; D001483:Base Sequence; D015342:DNA Probes / Q000235:genetics; D004279:DNA, Viral / Q000235:genetics; D004770:Enterovirus / Q000235:genetics / Q000302:isolation & purification*; D005516:Food Microbiology; D005602:France; D006507:Hepatovirus / Q000235:genetics / Q000302:isolation & purification*; D006801:Humans; D008969:Molecular Sequence Data; D016133:Polymerase Chain Reaction / Q000379:methods / Q000706:statistics & numerical data; D012367:RNA, Viral / Q000235:genetics / Q000302:isolation & purification; D012401:Rotavirus / Q000235:genetics / Q000302:isolation & purification*; D012680:Sensitivity and Specificity; D012758:Shellfish / Q000382:microbiology*; D014871:Water Microbiology
D015342:DNA Probes; D004279:DNA, Viral; D012367:RNA, Viral