Pig PCV2ORF2Protein Expression And The Establishment Of I-Elisa Detection Method And Pig Serum Antibody Detection And Analysis

Porcine circovirus type2(porcine circovirus2, PCV2) as the postweaning multisystemic wasting syndrome (postweaning multisystemic wasting syndrome, PMWS) pathogen, it can not only lead to the failure of weaning piglets, death, and the pig occurrence of multiple diseases closely related. Pigs infected with PCV2, the body will be in a state of immune suppression, and then cause various other disease pathogen of secondary infection, caused by PCV2indirect loss is estimated hard. PCV2at current has become severely hinder the healthy development of swine industry one of the main pathogens. At present, but also the lack of a suitable for large-scale diagnosis and detection of PCV2and reliable method, investigate its reason, it is PCV1in the current swine infection is quite common, and PCV1and PCV2nucleotide homology is very high in the detection of antigen cross phenomenon easily appeared. Because the ELISA method is very suitable for large-scale clinical samples, and PCV2specific antigen sites are concentrated in the ORF2area. If the clinical use PCV2ORF2to establish ELISA method for detection of the expression product, can achieve both effective to distinguish between PCV1and PCV2, and can carry out large-scale samples.In this research, the truncated ORF2gene of PCV2was cloned into the prokaryotic expressing vector pGEX-4T-1and expressed in the competent cells E.coli BL21(DE3) induced by the IPTG. Based on the purified inclusion bodies protein, an indirect ELISA method was established through pre-testing several parameters(temperature, IPTG concentration, inducing time, et al.).316sera submitted from four pig farms of shandong province were tested by the referred indirect ELISA. The experiment data were analyzed by the software SPSS v13.0. The result showed that the deviation was significant for these herds (P value<0.01), and the proportion of positive herds was90%(18/20), meanwhile remainder two from twenty farms were entirely negative. Among all sera, the positive rate was74.45%(235/316). It could be seen the seroprevalence specific for PCV2was in superior high degree according to these submitted sera from the four areas. And it is necessary that strengthening farm PCV2epidemic situation monitoring timely according to the planned vaccination.The results show that, this study to establish indirect ELISA for the detection of high sensitivity, strong specificity, and can be used for clinical PCV2serum antibody detection. This is a ELISA development of diagnostic kit development laid the foundation for the establishment of large scale pig farm, PCV2infection of rapid diagnosis method provides a scientific basis.