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.
MeSH terms
-
Animals
-
Base Sequence
-
Bivalvia / virology*
-
DNA Primers / genetics
-
Foodborne Diseases / prevention & control
-
Hepatitis A / prevention & control
-
Hepatovirus / genetics*
-
Hepatovirus / isolation & purification*
-
Hepatovirus / pathogenicity
-
Humans
-
Molecular Sequence Data
-
RNA, Viral / genetics*
-
RNA, Viral / isolation & purification*
-
RNA, Viral / standards
-
Reference Standards
-
Reproducibility of Results
-
Reverse Transcriptase Polymerase Chain Reaction / methods*
-
Reverse Transcriptase Polymerase Chain Reaction / standards
-
Reverse Transcriptase Polymerase Chain Reaction / statistics & numerical data
-
Shellfish / virology*