Sensitive And Specific Nest-PCR Detection Of The Two Major Shrimp Pathogenic Viruses (WSSV And IHHNV) Infection Of Penaeus Vannamei In HaiNan

Marine shrimp farming with its marked economic significance has long been taken into account as the major part of aquaculture industry. The demand of quality shrimp, which is among the top favorite seafood worldwide, is growing steadily each year in contrast to the depletion of natural shrimp stock in the wild occurring rapidly. However, disease caused by exotic viruses is the biggest single implement to aquaculture development and growth, in which outbreak of virus disease often cause devastating economic loss and impact on shrimp farming and industry. While early efforts to avoid shrimp disease were mainly relied on abandoning contaminated farms and expanding into clear water, practical and cost-effective means for the detection of shrimp pathogen viruses and management/limitation of their spread is the urgent focus of current scientific investigation.Han Nan, favored by its climate and abundance at its marine resources, has the leading role for shrimp farming and industry in Southwestern China. Penaeus vannamei is the major shrimp species aquacultured in Hainan. However, it is not yet clear about the identity and nature of viral pathogens that have caused shrimp virus disease in Hainan. To amplify SPF shrimp, we have to analyze and identify the kind of shrimp pathogen virus, and base on this, to establish a high sensitivity and specificity if this technique was applied to intense monitoring of the multiplying SPF shrimp.The object of our studies: To analyze and identify the causative virus species that are relevant to shrimp virus disease in Hainan. To develop and establish a highly sensitive and specific method for detection of the virus strains associated with infected shrimp in Hainan. The results of the ours studies: (1). WSSV and IHHNV shrimp virus are the major two types of virus that cause shrimp infection of Hai Nan aquaculture industry. Samples including Penaeus vannamei collected in the shrimp farms and others collected from a variety of markets in Haikou, Sanya, Danzhou, Qionghai, Wanning, Dongfang, and Wenchang have been analyzed by use of the virus detection method recommended by OIE. Nine strains of virus (WSSV, TSV, IHHNV), which has been observed in our country, have been found in these samples of which 67.6% contain WSSV, 45.9% contain IHHNV, 32.4% have both, and 5.4% contain TSV. (2) An approach by employing of multiplex nested-PCR assays has been established, which demonstrates satisfied sensitivity and specificity for convenient and rapid detection of WSSV and IHHNV viruses. Although PCR-based detection is cost- and labor-effective compared to the methods of nucleic acid hybridization, ELISA and others, the conventional PCR protocol in this regard is limited for detection of one virus strain a...