Colourimetric PCR-based detection of monodon baculovirus in whole Penaeus monodon postlarvae

The development of a nested polymerase chain reaction (PCR) assay is described to detect low concentrations of monodon baculovirus (MBV) DNA from total Penaeus monodon postlarval DNA. A modified DNA extraction procedure was also developed to circumvent problems associated with co-purification of PCR inhibitors in total DNA extracted from whole postlarvae, This method involved mechanical disruption of frozen prawn material immediately followed by phenol extraction at high temperature. An assessment of the sensitivity of the assay demonstrated detection down to eight viral genome equivalents. The PCR was shown to be specific for MBV DNA by not amplifying prawn DNA or DNA preparations of Baculovirus penaei (BP), white spot baculovirus (WSBV), bennettae baculovirus and insect Autographica californica nuclear polyhedrosis virus (NPV). A colourimetric method of PCR product detection was used to simplify final analysis. (C) 1998 Elsevier Science B.V. All rights reserved.