Multiplex RT-PCR assay for simultaneous detection of six viruses of penaeid shrimp

• Published in: Molecular and cellular probes 2008-06, Vol.22 (3), p.177-183
• Main Authors: Khawsak, Paisarn, Deesukon, Warin, Chaivisuthangkura, Parin, Sukhumsirichart, Wasana
• Format: Article
• Language: English
• Subjects: Animals; Baculovirus; DNA Primers - chemistry; DNA Viruses - genetics; DNA Viruses - isolation & purification; Hepatopancreatic parvo-like virus of shrimps; HPV; IHHNV; Infectious hypodermal and hematopoietic necrosis virus; MBV; Monodon; Multiplex RT-PCR; Parvovirus; Penaeidae - virology; Penaeus monodon; Reverse Transcriptase Polymerase Chain Reaction - methods; RNA Viruses - isolation & purification; RNA, Ribosomal - isolation & purification; Sensitivity and Specificity; Taura syndrome virus; TSV; White spot syndrome virus; WSSV; YHV
• ISSN: 0890-8508; 1096-1194
• DOI: 10.1016/j.mcp.2007.12.005

In the present study, multiplex reverse transcription-polymerase chain reaction (mRT-PCR) was developed for simultaneously detection of six major shrimp viruses including yellow-head virus (YHV), white spot syndrome virus (WSSV), Taura syndrome virus (TSV), hepatopancreatic parvovirus (HPV), infectious hypodermal and hematopoietic necrosis virus (IHHNV) and monodon baculovirus (MBV). The six primer sets could amplify viral nucleic acids resulting in PCR products with different sizes. They were highly specific and did not cross-hybridize with other viral or shrimp nucleic acids. The sensitivity of the multiplex RT-PCR was 0.15 pg for IHHNV, 0.15 pg for TSV, 1.00 pg for HPV, 1.5 pg for MBV, 5.00 pg for WSSV and 10.00 pg for YHV. In the field application, 42 samples including whole tissue of post-larvae and hepatopancreas of Penaeus monodon collected from ponds in the central and southern parts of Thailand during 2002–2005 were examined by multiplex RT-PCR. The results revealed that a single infection was dominant and WSSV was the highest prevalence at that time. Dual infection was found in one sample. This developed multiplex RT-PCR will be useful for simultaneous detection of six major viruses of penaeid shrimp and benefit to shrimp cultured industry.