A METHOD FOR NUCLEIC-ACID EXTRACTION FROM KURUMA PRAWN FOR PCR DETECTION OF PRDV (PENAEID ROD-SHAPED DNA VIRUS)

Penaeid rod-shaped DNA virus (PRDV) is the causative agent of penaeid acute viremia (PAV) in kuruma prawn (Penaeus japonicus) in Japan. To d etect the virus, a PCR method using ISOGEN (Nippongene) as a nucleic a cid extraction reagent has been used. In order to improve that method, an alternative extraction method named RNase-PEG precipitation (RNPP) method which includes phenol-chloroform extraction, RNase A treatment and polyethylene glycol precipitation was employed in this study. It was found that there was no significant difference in the amounts of e xtracted PRDV genome between the two methods. However, DNA preparation obtained by the ISOGEN method included contaminants in larger quantit ies than that by the present method. Comparison of the 2 PCR methods r evealed that the RNPP method had a higher sensitivity than the ISOGEN method in detecting PRDV.