肝胰腺细小病毒(HPV) PCR检测及流行情况调查

PCR Detection and Studies on the Prevalence of Hepatopancreatic Parvovirus (HPV)

中文关键词: 对虾  肝胰腺细小病毒(HPV)  PCR  检测

英文关键词: Penaeid shrimp  Hepatopancreatic parvovirus (HPV)  PCR  Detection

中文摘要:

      采用水产行业标准《对虾肝胰腺细小病毒病诊断规程 第1部分：PCR检测法》 (SC/T 7203.1- 2007)的方法，对2011−2013年期间我国沿海7个省市主要养殖对虾品种不同生长阶段的对虾样品进行该病毒携带情况的筛查。该方法的检测灵敏度为0.07 fg，相当于大约20个病毒拷贝。结果显示，639份样品的HPV阳性检出率为18.47%。其中，在中国对虾(Fenneropenaeus chinensis)、凡纳滨对虾(Litopenaeus vannamei)和日本囊对虾(Marsupenaeus japonicus)中均有阳性检出，且仔虾、幼虾、成虾各个阶段均可检出HPV，表明HPV已在我国养殖对虾中存在并流行。本研究结果为对虾养殖生产提供了疫病的科学数据，为我国养殖对虾中该病的流行情况提供了参考依据。

英文摘要:

      Hepatopancreatic parvovirus (HPV) is one of major pathogens responsible for shrimp diseases in the world. In recent years HPV disease has been found in coastal shrimp farming areas in China, however data about its prevalence has been unavailable. In this study, we examined shrimp samples at different developmental stages including egg, postlarvae, juvenile, adult and broodstock that were collected from the rearing ponds of seven coastal provinces in China (20112013). We detected the HPV in the samples using PCR according to the aquaculture industry standard “Diagnostic protocols for hepatopancreatic parvovirus disease of penaeid shrimp ——Part 1: PCR method” (SC/T 7203.1-2007). Our data showed that the primers were specific for HPV, and they had no cross-reaction with other pathogens such as infectious hypodermal and hematopoietic necrosis virus (IHHNV), white spot syndrome virus (WSSV), and specific pathogen free (SPF) shrimp DNA. Under the PCR conditions, the primers could detect as little as 0.07 fg purified HPV DNA which contained only 20 copies of HPV. This detection limit was 10-fold lower than that of real-time PCR that was established in our laboratory. Histopathological sectioning of HPV-infected Fenneropenaeus chinensis verified the existence of HPV inclusion bodies in the infected tissues. Under the optimized PCR conditions, a single product with the expected size (628 bp) was detected. The positive rates were 18.47% (118 out of 639) for penaeid shrimp, 20.36% for F. chinensis, 2.56% for Litopenaeus vannamei, and 50% for Marsupenaeus japonicus. The positive rates were 9.62% (5 out of 52), 20.28% (100 out of 493) and 100% (8 out of 8) for F.chinensis postlarvae, juvenile, and adult respectively. Furthermore, HPV was detected in penaeid shrimp samples at different developmental stages including postlarvae, juveniles and adults. Our study demonstrated that HPV has been prevalent in the farmed penaeid shrimp in China, and provided data about the shrimp disease and its epidemic situation in aquaculture industry. Future studies on the disease will help us understand its geographic distribution and host species, and help evaluate its prevalence.