| Feline calicivirus is a single-strand RNA viruses which one of Caliciviridae calicivirus genus members.The Virus diameter is 35-39 nm and no capsular. FCV can cause the cats’ mouth and upper respiratory disease. The disease is a multi-issue, epidemic, highly contagious infectious disease called infectious- rhinoconjunctivitis in the clinical. The infection animals do’nt express fatal disease, always show fever, rhinitis, stomatitis, increasing eye discharge. Aminals infected FCV alone does not lead fatal disease.When FCV infected with other pathogenic often show lameness, pneumonia, gastroenteritis, nephritis. Serious infection will caused cats dead. The disease is not only harmful to the cat of domesticated and the cat of tethering,but also threat to wild cats seriously. The best prevention is vaccination for FCV, but it can’t completely remove pathogens. the sick animal will became carriers of the virus source of infection detox virus to environment. Another,due to the high volatility of FCV,the immol/Lune protection will reduced.Due to FCV is extremely easy variability and the clinical symptoms diversity that brings some difficulties to disease detection and diagnosis. FCV can be detect by isolation of the virus, electron microscopy, RT-PCR, fluorescence quantitative PCR, and in the experiment, the agar diffusion. These methods can be carried out in ordinary laboratory,but need require skilled laboratory skills. Serological methods can either evaluate the immol/Lune protective effect of the vaccine, but also contribute to the clinical diagnosis,but currently there is no commol/Lercially available detection kit of serological.So it’s necessary that conducted serological diagnostic methods research for the prevention of disease control.FCV has three open reading frame(ORF), Which ORF2 encodes capsid protein that can produce antibodies effective for the body. capsid protein Contain multiple antigen recognition sites. In this study, we designed a pair of primers according to the gene sequences source feline calicivirus which stored in lab. We amplified 1 734 nucleotide sequences of ORF2,which cloned into the expression vector p ET-28 a. that we constructed recombinant plasmid p ET-28a-FCV1734 successfully. The plasmid was transformed into E. coli expression strain Detect the induced expression products by SDS-PAGE and Western blot, proved the expressed protein have a good reactogenicity. purified and extract the protein, as a feline calicivirus antigen of indirect ELISA method. Use square matrix method to determine the optimal working concentration secondary antibody-HRP was 1: 5000,the best antigen coating concentration was 2.1250μg/m L, The best anti-dilution was 1: 200. Optimized the optimal reaction conditions. Determine the the shade,and the positive critical value,when OD490 value greater than 0.2570 was determined to be positive, OD490 value less than 0.2390 was determined to be negative, OD490 between 0.2570-0.2390 was determined suspected. Using feline calicivirus indirect ELISA method detected the cat serum 49 parts from Jilin Province,188 parts from Shanghai and 47 parts from Guangxi Province to established in this study, the results show that the seroprevalence of Jilin cats was 69.39%(34/49), the seroprevalence of Shanghai cats was 91.49%( 172/188) and the seroprevalence of Guangxi cats was 85.92%(244/288)。... |
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