Key result
A competitive RT-PCR method with coextraction of standard RNA enabled reproducible quantification of Hepatitis A virus in shellfish, with a quantifiable range of 10^4 to 10^7 copies per gram.
A competitive RT-PCR method with an internal standard RNA allows for reproducible quantification of hepatitis A virus in contaminated shellfish.
Enables reproducible HAV quantification in shellfish; leaves open validation for routine food safety surveillance.
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.
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Arnal et al. (1999) studied Hepatitis A Virus contamination in shellfish. Competitive RT-PCR with internal standard RNA was evaluated on Quantifiable range of HAV. A competitive RT-PCR method with coextraction of standard RNA enabled reproducible quantification of Hepatitis A virus in shellfish, with a quantifiable range of 10^4 to 10^7 copies per gram.
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