Detection of penaeid rod-shaped DNA virus (PRDV) by PCR

We have developed a method for detection of penaeid rod-shaped DNA virus (PRDV, formerly RV-PJ), the causative agent of penaeid acute viremia (PAV) of kuruma shrimp, by polymerase chain reaction (PCR). Two pairs of PCR primers (P1/P2 and P3/P4) were prepared, based on the nucleotide sequence of the PRDV genome. PCR using either P1/P2 or P3/P4 specifically amplified PRDV DNA. When PCR products amplified with P1/P2 were subjected to nested PCR with P3/P4, a higher sensitivity was seen than with PCR using P1/P2 or P3/P4 alone. The nested PCR was capable of detecting PRDV from a symptomatic kuruma shrimp that had been experimentally infected with low doses of PRDV, excretions from experimentally infected kuruma shrimp, and other crustaceans in a culture pond where spontaneous PAV occurred.