多重RT-PCR方法在凡纳滨对虾病毒检测中的应用

Multiplex RT-PCR Assay for Detection of Viruses of Invalid Penaeus vannamei in Guangxi

病毒检测在预防和控制对虾病毒性疫病实践中具有重要的作用和意义。为了提高对虾病毒检测的快速性和简便性,建立了一种可同时检测6种对虾病毒的RT-PCR方法,并利用该方法对来自广西沿海地区的130份病虾样品中的黄头病毒(YHV)、白斑综合征病毒(W SSV)、桃拉病毒(TSV)、肝胰腺坏死病毒(HPV)、传染性皮下组织坏死病毒(IHHNV)以及杆状病毒(MBV)进行了检测。检测结果:白斑综合征病毒发病率最高,检出率高达73.8%;传染性皮下组织坏死病毒发病率为18.5%;桃拉病毒检出率为3.1%;未检出黄头病毒、肝胰腺坏死病毒和杆状病毒。该RT-PCR方法扩增得到的片段经测序,结果与标准毒株的核酸数据高度吻合。实践证明,该实验建立的RT-PCR方法具有高度的准确性,与其他检测方法相比具有快速、简便的特点,在对虾病毒检疫、流行病学调查等方面具有重要的应用价值。

Viral detection was important and useful to protect the shrimp against virus infection. In order to improve the viral detection method in shrimp , a more rapid and convenient multiplex reverse transcfiption-polymerase chain reaction （mRT-PCR） was established in the present study. The new method was used for detection of shrimp viruses, including Yellow-head virus（YHV） ,white spot syndrome virus （WSSV）, Tanra syndrome virus （TSV） , hepatopancreatic parvovirus （HPV） , infectious hypodermal and hematopoietic necrosis virus （IHHNV） and monodon baculovirus （MBV）. 130 ill shrimp samples of Penaeus vannamei were collected from the coastal areas in Guangxi . The detect result revealed that WSSV infection rate was 73.8 % , IHHNV infection rate was 18.5 % ,TSV infection rate was 3.1% . none of YHV, HPV and MBV was detected in these samples . The sequence result of amplify bands showed highly homology with the nucleic data of standard virus strains . Comparing to other detection methods,this new mRT-PCR method was not only precise but more rapid and convenient, and it may be useful in the quarantine and prevalent investigation to shrimp Penaeus varmamei cultured industry.