Development of a PCR procedure for the detection of Baculovirus penaei in shrimp

Abstract

A PCR-based detection procedure for Baculovirus penaei (BP) was developed. Three forward and 3 reverse primers were designed to provide 6 primer-pair combinations with PCR products ranging in size from 196 to 933 bp. The expected amplification products were obtained when template DNA isolated from postiarval Penaeus vannamei experimentally infected with BP in the laboratory or from wild-caught BP-infected juvenile P. aztecus were used. DNA isolated from specific pathogen-free P. vannamei did not yield amplification products using the same PCR primers. The procedure used to prepare the sample for PCR was found to be critical because of the presence of unknown compounds in shrimp tissue that inhibited DNA polymerase. A suitable sample preparation procedure using proteinase K digestion, phenol/chloroform extraction and ethanol precipitation is described.