Polymerase chain reaction (pCR)-based rapid identification of white spot syndrome virus in shrimp, Penaeus monodon

Abstract

The incidence of white spot syndrome virus (WSSV) in Penaeus monodon from the coastal region of Bangladesh was determined by a two-step polymerase chain reaction (PCR)-based screening protocol using two pairs of oligonucleotide primers directed towards WSSV DNA. The widespread occurrence of WSSV and the effectiveness of the protocol were further ensured by investigating with horina chingri, Metapenaeus monoceros and mud crab, Scylla serrata. The experiment was conducted with tissue samples collected from muscle, hepatopancreas and pleopod together with individuals from two different farms; apparently disease infected farm and uninfected farm, situated in the same area. The first-step PCR of total genomic DNA isolated from the tissue samples served as the templates for the second-step PCR, which resulted in DNA amplification of an expected size for each sample. The easiness, cost-effectiveness and high specificity of this method rely on using a single-step DNA preparation without involving hazardous organic solvents as well as two pairs of oligonucleotide primers highly specific for WSSV gene. This is the first scientific report on applying PCR-based WSSV screening protocol in Bangladesh without using patented WSSV detection kits as in elsewhere in the country. The pioneering works in Bangladesh and the results suggest that a PCR screening of WSSV infection for shrimp brood and larval stocks and other potential virus carriers could be an effective. tool in fight against the deadly virus that has almost devastated the shrimp industry of the country.