title: Quantification of hepatitis A virus in shellfish by competitive reverse transcription-PCR with coextraction of standard RNA.
abstract: To quantify hepatitis A virus (HAV) in experimentally contaminated mussels, we developed an internal standard RNA with a 7-nucleotide deletion for competitive reverse transcription (RT)-PCR. Deposited directly into the sample, this standard was used both as extraction control and as quantification tool. After coextraction and competitive RT-PCR, standard and wild-type products were detected by differential hybridization with specific probes and a DNA enzyme immunoassay. The quantifiable range with this reproducible method was 10(4) to 10(7) copies of HAV/gram or 400 to 10(6) 50% tissue culture infective doses/ml.
authors.affiliation: Laboratoire de Virologie, Institut de Biologie, Centre Hospitalier Universitaire, 44093 Nantes Cedex, France. sbi@sante.univ-nantes.fr
authors.forename: C
authors.initials: C
authors.lastname: Arnal
authors.forename: V
authors.initials: V
authors.lastname: Ferre-Aubineau
authors.forename: B
authors.initials: B
authors.lastname: Mignotte
authors.forename: B M
authors.initials: BM
authors.lastname: Imbert-Marcille
authors.forename: S
authors.initials: S
authors.lastname: Billaudel
mesh_terms: D000818:Animals; D001483:Base Sequence; D049872:Bivalvia / Q000821:virology*; D017931:DNA Primers / Q000235:genetics; D005517:Foodborne Diseases / Q000517:prevention & control; D006506:Hepatitis A / Q000517:prevention & control; D006507:Hepatovirus / Q000235:genetics* / Q000302:isolation & purification* / Q000472:pathogenicity; D006801:Humans; D008969:Molecular Sequence Data; D012367:RNA, Viral / Q000235:genetics* / Q000302:isolation & purification* / Q000592:standards; D012015:Reference Standards; D015203:Reproducibility of Results; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000379:methods* / Q000592:standards / Q000706:statistics & numerical data; D012758:Shellfish / Q000821:virology*
chemical_list: D017931:DNA Primers; D012367:RNA, Viral