Using a pair of specific primers designed according to the relevant nucleotide sequences from GenBank, the main antigen domain for VP2 gene of Porcine parvovirus was ampilified with PCR method using the genomic DNA as...Using a pair of specific primers designed according to the relevant nucleotide sequences from GenBank, the main antigen domain for VP2 gene of Porcine parvovirus was ampilified with PCR method using the genomic DNA as template. The PCR product was cloned into the expression vector pIREShyg to get a recombinant eukaryotic expression plasmid pIREShyg-VP2, which was then transfected into the CHO-K1 cells. The expressed product was detected by IFA after the positive cell clone was selected with hygromycin. The result revealed that the main antigen domain for VP2 gene of porcine parvovirus was stably expressed in CHO-K1 cells.展开更多
The purified recombinant nuclocapsid protein of PRRSV was used to conjoined with latex, and the conjoined concentration,temperature and time were optimized, a detection method for serum antibodies against Porcine repr...The purified recombinant nuclocapsid protein of PRRSV was used to conjoined with latex, and the conjoined concentration,temperature and time were optimized, a detection method for serum antibodies against Porcine reproductive and respiratory syndrome virus was established. Some tests for the method’s sensitivity, specificity and stability were confirmed. About 200 serum samples were detected by using the method, and these samples were also detected by recombinant N protein based ELISA and IDEXX ELISA kit. The result showed that the agreement ratio of the recombinant N protein based LAT with other two methods arrived at 93%, 78%, respectively.展开更多
文摘Using a pair of specific primers designed according to the relevant nucleotide sequences from GenBank, the main antigen domain for VP2 gene of Porcine parvovirus was ampilified with PCR method using the genomic DNA as template. The PCR product was cloned into the expression vector pIREShyg to get a recombinant eukaryotic expression plasmid pIREShyg-VP2, which was then transfected into the CHO-K1 cells. The expressed product was detected by IFA after the positive cell clone was selected with hygromycin. The result revealed that the main antigen domain for VP2 gene of porcine parvovirus was stably expressed in CHO-K1 cells.
文摘The purified recombinant nuclocapsid protein of PRRSV was used to conjoined with latex, and the conjoined concentration,temperature and time were optimized, a detection method for serum antibodies against Porcine reproductive and respiratory syndrome virus was established. Some tests for the method’s sensitivity, specificity and stability were confirmed. About 200 serum samples were detected by using the method, and these samples were also detected by recombinant N protein based ELISA and IDEXX ELISA kit. The result showed that the agreement ratio of the recombinant N protein based LAT with other two methods arrived at 93%, 78%, respectively.