Comparison of different primer sets for the RT-PCR detection of hepatitis A virus and astrovirus in mussel tissues.
In the present study, the efficiency of several primer sets for the RT-PCR detection of hepatitis A virus (HAV) and astrovirus from both crude viral extracts and experimentally infected shellfish tissues was evaluated. Differences were observed depending on the primer set employed in the sensitivity of amplification of both viral types. For HAV primers, HAV240/HAV68 yielded the higher sensitivity: showing a detection limit of 0.02-0.1 infectious particles/microL or mg of tissue (either crude extracts or seeded mussel tissues). Regarding detection of AsV, a better performance was observed with primer set A1/A2 achieving a sensitivity of 0.1-1 PFU/microL or mg of tissue. The results obtained in this work strongly indicated that selection of primer sets to be employed for the routine detection of enteric viruses was a critical point in the design of the RT-PCR protocols.
Romalde
J L
JL
Dept de Microbiología y Parasitologia, Facultad de Biología, Universidad de Santiago de Compostela 15782, Spain. mpromald@usc.es
Ribao
C
C
Luz Vilariño
M
M
Barja
J L
JL
D000818:Animals; D019200:Astroviridae / Q000235:genetics*; D049872:Bivalvia / Q000821:virology*; D017931:DNA Primers; D005506:Food Contamination; D006506:Hepatitis A / Q000235:genetics*; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000379:methods*
D017931:DNA Primers