Study On The Detection Of WSSV And IHHNV In Shrimp Using Real-Time Quantitative PCR

Shrimp cultural is one of the aquiculture pillar industries, It's also a major exporting aquatic product. Since the increasingly severe viral diseases, most of shrimp cultural regions are suffering great economic losses. A brief review on recent advances in shrimp viral pathogens, and new techniques for the diagnosis of shrimp virus. According to White spot syndrome virus (WSSV) and Infections Hypodermal and Haematopoietic Nerosis Viurs (IHHNV) exsiting in shrimp cultural industry, The study established a TaqMan-based multiplex real-time PCR assay for detection of WSSV and IHHNV. Some clinical samples collected from several aquatic product markets in Xiamen, Fujian, PRC were detected by real-time PCR assay. It was aimed at establishing a quick,simple,sensitivite and specific tool for laboratory of quarantine department. The results are as follows:1 .A mutiplex PCR was optimized to detect two pathogens,WSSV and IHHNV, in shrimp. Two pairs of specific primers were designed according to the conserved regions. It demonstrated that this multiplex PCR could be used as a sensitive and quick tool to detect WSSV and IHHNV in clinical samples simultaneously.2.Fluorescent Quantitive Polymerase Chain Reaction(FQ-PCR) assay for detection of WSSV and IHHNV were established respectively. The PCR conditions were optimzed to improve the sensitivity, specifity and repeatability. A standard curve was constructed and the result showed that the sensitivity of the method was 10 copies/μl and the linear relation was fitted.3. To our knowledgement, this is the first report for real-time PCR assay for the simultaneous detection of WSSV and IHNNV. The sequence downloaded from Genbank was aligned by biological software and the specific primers and probes were designed in the conserved region of WSSV. The PCR conditions were optimzed to improve the sensitivity, specifity and repeatability. The relative quantification in real-time PCR to detection of WSSV and IHNNV were establised with this method, and the good results were achieved by using the method in the actual application.