Development of a polymerase chain reaction (PCR) procedure for the detection of baculovirus associated with white spot syndrome (WSBV) in penaeid shrimp

摘要

The causative viral agent was purified from diseased shrimp (Penaeus japonicus) with WSBV. Several hundred clones were obtained from libraries of the purified viral genomic DNA. Results from nucleotide sequence analysis showed none of the WSBV clones had considerable sequence homology with those of other known viruses. Using the sequence data of WSBV genomic DNA, a pair of PCR primers was designed. After 30 cycles of PCR amplification of viral genomic DNA extracted from WSBV, a single product of the expected size was detected. Southern blot hybridization confirmed that the amplified product was specific to the DNA of WSBV. The PCR system was able to detect 1 pg of WSBV DNA after 30 cycles and efficiently amplify the target region of the WSBV genein the nucleic acids extracted either from the diseased shrimps or hatchery shrimps with no signs of viral infection.