Established IELISA Of Type1and The Serological Survey Of Duck Circovirus

Duck circovirus (DuCV) is listed as a member of the circovirus of the family Circoviridae, which is a single-stranded circular DNA virus without envlope. Since2003DuCV firstly was descovered in Germany, it was found and reported in Hungary. United States,TaiWan and China. In this study, we established iELISA for the genotype1of DuCV to study the prevalent of duck circovirus (DuCV) in Shandong province, a total of1130sera collecting from Weifang and Taian regions. The research contains three parts:1. Development of Indirect ELISA for Detection of Antibodies to Genotype1of DuCV and Preliminary ApplicationThe study established iELISA for detecting the antibodies of genotype1of DuCV. The partial Cap genotype1of Duck circovirus (DuCV)was amplified by polymerase chain reaction(PCR) and and then cloned in prokaryotic vector pET-32a (+) vector to construct the recombinant plasmid. After the induction of IPTG, the purified recombinant protein can be antigen to found an iELISA to detect the antibody of DuCV. The results indicated that:The cap genotype1of DuCV can expressed in E.coli efficiently and steadily.Western blot analysis proved the renaturation protein has good immunoreactivity against DuCV positive antibody. The indirect ELISA method for the detection of DuCV specific antibody in duck serum was established, after the optional working circumstances for the ELISA assay (antigen concentration:800ng/well; optimal serum dilution:1:50; positive Cut-off value:0.324with chessboard titration. The results showed that the individual positive ratio is4%～78.125%in healthy duck groups,33.80%(382/1130) in average,100%(colonial) in duck flocks.2. Serological Survey of Duck Circovirus in Parts of Shandong Province in2011In order to study the prevalent of the type2of duck circovirus (DuCV) in Shandong province, a total of1130sera collected from Weifang and Taian regions were tested by indirect ELISA for detection of antibodies to DuCV. The results indicated that DuCV-specific antibodies were detected in338(29.91%) of1130samples obtained from17meat Cherry Valley duck flocks. The positive rates for each of the17flocks ranged from2%to78%. The DuCV-positive rate of42.81%in Weifang region was significantly higher than that in Taian region (17.73%), indicating the higher prevalence of DuCV in duck flocks of long-term fanned region.3. Coinfection Analysis of Genotype1and2of DuCVIt was analysed that the result of serological survey of the duck flocks in Weifang and Taian in Shandong province detected with iELISA for detecting genotype1and genotype2of DuCV. The results showed that antibody positive coincidence rate of genotype1and genotype2was80.7%。Both genotypes were positive is63in Taian, accounting for10.84%of the total number of detection. According to Weifang, it was slightly higher than Taian as10.84%. The coinfection positive rate of genotype1and2of DuCV was16.99%, which indicated that the ducks were seriously coinfected by the two genotypes of DuCV in Shandong province.