A comparison of RT-PCR-based assays for the detection of HAV from shellfish.
The hepatitis A virus (HAV) is the most common cause of viral infection linked to shellfish consumption. The lack of correlation between the fecal coliform indicators and the presence of enteric viruses in shellfish and their harvesting waters points to the need for molecular methods to detect viruses. We compared two RT-PCR based techniques currently available for the detection of the hepatitis A virus (HAV) in shellfish. Both approaches involve extraction of viral particles by glycine buffer and concentration of virus particles by one or two PEG precipitation steps. One procedure involves as RNA extraction method the use of oligo (dT) cellulose to select poly (A) RNA, and the other uses a system in which total RNA is bound on silica membrane. Comparison of the two RT-PCR based methods highlighted the efficiency of the first approach which is less time-consuming and technically demanding than the second.
Di Pinto
A
A
Dipartimento di Sanità e Benessere degli Animali, Università degli Studi, Bari, Italy.
Conversano
M C
MC
Forte
V T
VT
La Salandra
G
G
Montervino
C
C
Tantillo
G M
GM
D000818:Animals; D049872:Bivalvia / Q000821:virology*; D006506:Hepatitis A / Q000517:prevention & control* / Q000635:transmission; D030041:Hepatitis A virus / Q000235:genetics* / Q000302:isolation & purification; D008567:Membranes, Artificial; D012333:RNA, Messenger / Q000032:analysis / Q000302:isolation & purification; D012367:RNA, Viral / Q000032:analysis / Q000302:isolation & purification; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000379:methods*; D012680:Sensitivity and Specificity; D012758:Shellfish / Q000821:virology*; D012822:Silicon Dioxide
D008567:Membranes, Artificial; D012333:RNA, Messenger; D012367:RNA, Viral; D012822:Silicon Dioxide