| Summary: | 碩士 === 國立屏東科技大學 === 動物疫苗科技研究所 === 104 === Koi herpesvirus (Cyprinid herpesvirus 3, CyHV-3) is a large double-stranded (ds) DNA virus of fish that has been assigned to the family Alloherpesviridae within the order Herpesvirales , CyHV-3 causes mass mortalities and huge economic losses to the carp aquaculture industry. The goal of this study is to develop an inactivated vaccine and an optimal immunization program for koi . We establish primary cells derived from koi tissues and inoculated koi cells with KHV-infected fish tissue under the optimum culture conditions and subsequently isolated KHV. In addition,TCID50 and quantitative polymerase chain reaction (qPCR) assay were developed using SYBR Green to detect and quantify koi herpesvirus in KHV-inoculated cell or naturally infected koi. Finally, we will perform animal experiments to evaluate the protective efficacy of the inactivated vaccines. The isolate of KHV was successfully passaged in koi brain cells. Titer of the virus gradually increased, reaching 108 TCID50/ ml at passage 25.When 107 TCID50/ ml of this isolate was inactivated and used to immunize fish, the neutralization titer, reached 32x dilution 42 days after immunization. Our study showed that this strain retains high antigencity and can elicit high neutralizing antibody titer, thus , could be further developed as an inactivated vaccine.
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