VP1, a capsid protein of swine vesicular disease virus, was cloned from the SVDV HK/70 strain and inserted into retroviral vector pBABE puro, and expressed in PK15 cells by an retroviral expression system. The ability...VP1, a capsid protein of swine vesicular disease virus, was cloned from the SVDV HK/70 strain and inserted into retroviral vector pBABE puro, and expressed in PK15 cells by an retroviral expression system. The ability of the VP1 protein to induce an immune response was then evaluated in guinea pigs. Western blot and ELISA results indicated that the VP1 protein can be recognized by SVDV positive serum, Furthermore, anti-SVDV specific antibodies and lymphocyte proliferation were elicited and increased by VP1 protein after vaccination. These results encourage further work towards the development of a vaccine against SVDV infection.展开更多
The capsid protein precursor （P1）, which plays a major role for the generation of polypeptides of swine vesicular disease virus （SVDV）, was cloned from SVDV HK/70 strain into the retroviral vector pBABE puro and e...The capsid protein precursor （P1）, which plays a major role for the generation of polypeptides of swine vesicular disease virus （SVDV）, was cloned from SVDV HK/70 strain into the retroviral vector pBABE puro and expressed in the mammalian cell line PK15 through the retroviral expression system. The activity of recombinant protein to induce immune response was evaluated in guinea pigs. IFA and Western Blot were used to detect the recombinant protein expression. The results showed that the recombinant protein could be recognized by SVDV positive serum, and animal test showed SVDV-specific antibodies. All of those results indicate that a retroviral-based vaccine carrying the capsid protein precursor （P1） of SVD is able to be expressed in the eukaryotic cell and elicites strong SVDV-specific immune responses in guinea pigs.展开更多
[Objective] The aim of this study was to develop an indirect sandwich EUSA method for detection of swine vesicular disease virus （SVDV）. [Method] High titer serum against SVDV was prepared respectively by inoculated...[Objective] The aim of this study was to develop an indirect sandwich EUSA method for detection of swine vesicular disease virus （SVDV）. [Method] High titer serum against SVDV was prepared respectively by inoculated rabbits and guinea pigs with purified virus. To develop an indirect sandwich ELISA, the optimum concentrations of capture antibody, detection antibody, enzyme conjugate and standard antigen were determined using block titration, and positive threshold value was also determined. The specificity, sensitivity and repeatability of the developed IELISA were evaluated using cross-reaction test, comparison test and intra-assay repeated test. In addition, standard samples and clinical samples were detected by this method. [ Result] The best working conditions of the developed ELISA are as follows： capture antibody, 1：400; detection antibody, 1 ： 200; enzyme conjugate, 1 ： 8 000; and standard antigen, 1 ： 4. The positive threshold value was found to be 0.20. For the detection by the developed EUSA, no cross-reaction with foot and mouth disease was observed. The developed ELISA had close sensitivity with ELISA recommended by the World Organisation for Animal Health （OIE） but had sensitivity 2 -4 times higher than that of reverse indirect hernagglutination test. In addition, the developed method also had good reproducibility, and the detection results of standard samples were in line withtheir own background. All the 36 clinical samples were negative in the developed ELISA. [ Conclusion] The developed indirect sandwich ELISA can be used for diagnosis of swine vesicular disease.展开更多
The full-length cDNA clone of swine vesicular disease virus HK/70 strain named pSVOK12 was constructed in order to study the antigenicity, replication, maturation and pathogenicity of swine vesicular disease virus. In...The full-length cDNA clone of swine vesicular disease virus HK/70 strain named pSVOK12 was constructed in order to study the antigenicity, replication, maturation and pathogenicity of swine vesicular disease virus. In vitro transcription RNA from pSVOK12 transfected IBRS-2 cells and the re- covered virus RNA were isolated and sequenced, then indirect hemagglutination test, indirect im- munofluorescence assays, eleectron microscope test, 50% tissue culture infecting dose (TCID50) assays and mouse virulence studies were performed to study the antigenicity and virulence of the recovered virus. The result showed that the infectious clones we ob- tained and the virus derived from pSVOK12 had the same biological properties as the parental strain HK/70. The full-length infectious cDNA clone, pSVOK12, will be very useful in studies of the anti- genicity, virulence, pathogenesis, maturation and replication of SVDV.展开更多
文摘VP1, a capsid protein of swine vesicular disease virus, was cloned from the SVDV HK/70 strain and inserted into retroviral vector pBABE puro, and expressed in PK15 cells by an retroviral expression system. The ability of the VP1 protein to induce an immune response was then evaluated in guinea pigs. Western blot and ELISA results indicated that the VP1 protein can be recognized by SVDV positive serum, Furthermore, anti-SVDV specific antibodies and lymphocyte proliferation were elicited and increased by VP1 protein after vaccination. These results encourage further work towards the development of a vaccine against SVDV infection.
基金Supported by Key Technology R&D Programme (2006BAD06A03)
文摘The capsid protein precursor （P1）, which plays a major role for the generation of polypeptides of swine vesicular disease virus （SVDV）, was cloned from SVDV HK/70 strain into the retroviral vector pBABE puro and expressed in the mammalian cell line PK15 through the retroviral expression system. The activity of recombinant protein to induce immune response was evaluated in guinea pigs. IFA and Western Blot were used to detect the recombinant protein expression. The results showed that the recombinant protein could be recognized by SVDV positive serum, and animal test showed SVDV-specific antibodies. All of those results indicate that a retroviral-based vaccine carrying the capsid protein precursor （P1） of SVD is able to be expressed in the eukaryotic cell and elicites strong SVDV-specific immune responses in guinea pigs.
基金funded by the Major Sci-Tech Fund of Gansu Province (092NKDA032)
文摘[Objective] The aim of this study was to develop an indirect sandwich EUSA method for detection of swine vesicular disease virus （SVDV）. [Method] High titer serum against SVDV was prepared respectively by inoculated rabbits and guinea pigs with purified virus. To develop an indirect sandwich ELISA, the optimum concentrations of capture antibody, detection antibody, enzyme conjugate and standard antigen were determined using block titration, and positive threshold value was also determined. The specificity, sensitivity and repeatability of the developed IELISA were evaluated using cross-reaction test, comparison test and intra-assay repeated test. In addition, standard samples and clinical samples were detected by this method. [ Result] The best working conditions of the developed ELISA are as follows： capture antibody, 1：400; detection antibody, 1 ： 200; enzyme conjugate, 1 ： 8 000; and standard antigen, 1 ： 4. The positive threshold value was found to be 0.20. For the detection by the developed EUSA, no cross-reaction with foot and mouth disease was observed. The developed ELISA had close sensitivity with ELISA recommended by the World Organisation for Animal Health （OIE） but had sensitivity 2 -4 times higher than that of reverse indirect hernagglutination test. In addition, the developed method also had good reproducibility, and the detection results of standard samples were in line withtheir own background. All the 36 clinical samples were negative in the developed ELISA. [ Conclusion] The developed indirect sandwich ELISA can be used for diagnosis of swine vesicular disease.
文摘The full-length cDNA clone of swine vesicular disease virus HK/70 strain named pSVOK12 was constructed in order to study the antigenicity, replication, maturation and pathogenicity of swine vesicular disease virus. In vitro transcription RNA from pSVOK12 transfected IBRS-2 cells and the re- covered virus RNA were isolated and sequenced, then indirect hemagglutination test, indirect im- munofluorescence assays, eleectron microscope test, 50% tissue culture infecting dose (TCID50) assays and mouse virulence studies were performed to study the antigenicity and virulence of the recovered virus. The result showed that the infectious clones we ob- tained and the virus derived from pSVOK12 had the same biological properties as the parental strain HK/70. The full-length infectious cDNA clone, pSVOK12, will be very useful in studies of the anti- genicity, virulence, pathogenesis, maturation and replication of SVDV.
