A Method for Nucleic Acid Extraction from Kuruma Prawn for PCR Detection of PRDV (Penaeid Rod-shaped DNA Virus)

Abstract

Penaeid rod-shaped DNA virus (PRDV) is the causative agent of penaeid acute viremia (PAV) in kuruma prawn (Penaeus japonicus) in Japan. To detect the virus, a PCR method using ISOGEN (Nippongene) as a nucleic acid 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 extracted PRDV genome between the two methods. However, DNA preparation obtained by the ISOGEN method included contaminants in larger quantities than that by the present method. Comparison of the 2 PCR methods revealed that the RNPP method had a higher sensitivity than the ISOGEN method in detecting PRDV.