二温式PCR检测对虾白斑综合征病毒

Development and applification of two-temperature PCR for the detection of white spot syndrome virus in penaeid shrimp

本研究设计了一对能扩增大小为 30 6 bp对虾白斑综合征病毒 (WSSV)某段基因的特异性引物 ,优化建立了能快速检测 WSSV的二温式 PCR。在对包括 10 5份临床样品、10份 SPF南美白对虾组织样品和其他对虾病害病原在内的样品检测结果中 ,有 6 5份临床样品呈现 WSSV阳性 ,而 10份 SPF南美白对虾组织样品和其他对虾病害病原的 PCR结果为阴性。该二温式 PCR最低能检测到 1pg的 WSSV感染对虾组织样品总 DNA。这些结果表明 ,该 PCR具有高度的特异性和敏染性。

A two-temperature PCR which can amplify 306 base pair of the genome of white spot syndrome virus (WSSV) in penaeid shrimp was developed and optimized in this study.The test of sensitivity of this PCR showed that it could detected as low as 1pg of WSSV total DNA.We used this PCR as a diagnostic method to detect WSSV in the cultivated penaeid shrimps in Guangxi.Results showed that 65 of 105 clinical penaeid shrimp samples were found positive for WSSV and negative for the controls,including tissues from a pathogen-free P.vannamei shrimp sample from Supper Shrimp Inc.and other pathogens identified and saving at our lad, such as Taura syndrome virus(TSV), yellowhead virus(YHV),infectious hypodermal and haematopoietic necrosis virus(IHHNV) or Vibrio cholerae(V.Cholerae). The results demonstrated that this two-temperature PCR can be used as a rapid and sensitive detection method for WSSV diagnosis.