Simultaneous PCR detection of two shrimp viruses (WSSV and MBV) in postlarvae of Penaeus monodon in the Philippines

Abstract

A duplex polymerase chain reaction (PCR) protocol was developed for the simultaneous detection of two penaeid shrimp viruses, namely, White spot syndrome virus (WSSV) and Monodon baculovirus (MBV) infecting Penaeus monodon in Philippines. The method was designed for screening postlarval samples with dual infections of MBV and WSSV. The developed protocol was able to generate a 211 bp amplicon which is highly specific for WSSV and a 361 bp amplicon specific for MBV. An assessment of the sensitivity of the developed duplex PCR demonstrated the detection of both the amplicons up to 0.1 femtogram (fg) of plasmid DNA containing the target sequences equivalent to 15 copies of the viral target sequence. In addition to its high specificity and sensitivity, the developed duplex PCR offers an efficient and rapid tool for screening penaeid shrimp viruses since both WSSV and MBV can be diagnosed in a single reaction.