A sandwich format immunochromatographic assay for detecting foot-and-mouth disease virus (FMDV) serotypes was developed. In this rapid test,affinity purified polyclonal antibodies from Guinea pigs which were immunized...A sandwich format immunochromatographic assay for detecting foot-and-mouth disease virus (FMDV) serotypes was developed. In this rapid test,affinity purified polyclonal antibodies from Guinea pigs which were immunized with sucking-mouse adapted FMD virus (A/AV88(L) strain) were conjugated to colloidal gold beads and used as the capture antibody,and affinity purified polyclonal antibodies from rabbits which were immunized with cell-culture adapted FMD virus (A/CHA/09 strain) were used as detector antibody. On the nitrocellulose membrane of the immunochromatographic strip,the capture antibody was laid on a sample pad,the detector antibody was printed at the test line(T) and goat anti-guinea pigs IgG antibodies were immobilized to the control line(C). The lower detection limit of the test for a FMDV 146S antigen is 11.7ng/ml as determined in serial tests after the strip device was assembled and the assay condition optimization. No cross reactions were found with FMDV serotype C,Swine vesicular disease (SVD),Vesicular stomatiti svirus (VSV) and vesicular exanthema of swine virus (VES) viral antigens with this rapid test. Clinically,the diagnostic sensitivity of this test for FMDV serotypes A was 88.7% which is as same as an indirect-sandwich ELISA. The specificity of this strip test was 98.2% and is comparable to the 98.7% obtained with indirect-sandwich ELISA. This rapid strip test is simple,easy and fast for clinical testing on field sites; no special instruments and skills are required,and the result can be obtained within 15 min. To our knowledge,this is the first rapid immunochromatogarpic assay for serotype A of FMDV.展开更多
The peptide of amino acids 141～160 of VP1 protein of foot\|and\|mouth disease virus (FMDV) is a major B cell epitope and the peptide of amino acids 21～40 is an important T cell epitope.In this study,the DNA fragment...The peptide of amino acids 141～160 of VP1 protein of foot\|and\|mouth disease virus (FMDV) is a major B cell epitope and the peptide of amino acids 21～40 is an important T cell epitope.In this study,the DNA fragments of 141～160 and 21～40 peptide epitopes of a strain of type O FMDV was chemically synthesized and arranged into a tandem repeat 141～160 (20AA)\|21～40 (20AA)\|141～160 (20AA).This tandem sequence was fused to the 3′ end of the heavy chain constant region gene of swine immunoglobulin G and was then cloned into mammalian expression vector pCDM8 to form a recombinant plasmid pCDM8FZ3.After pCDM8FZ3 was inoculated intramuscularly into guinea pigs,it elicited a neutralizing antibody response and a specific spleen T cell proliferative response,and 66% of the vaccinated animals were protected from viral challenge.Our study indicated that the heavy chain constant region of swine IgG can act as the carrier protein for FMDV peptide epitopes,and pCDM8FZ3 is a potential DNA vaccine candidate to prevent FMDV infection.展开更多
基金Financial supported by the Gansu ProvincialSci. & Tech. Department (1002NKDA037)
文摘A sandwich format immunochromatographic assay for detecting foot-and-mouth disease virus (FMDV) serotypes was developed. In this rapid test,affinity purified polyclonal antibodies from Guinea pigs which were immunized with sucking-mouse adapted FMD virus (A/AV88(L) strain) were conjugated to colloidal gold beads and used as the capture antibody,and affinity purified polyclonal antibodies from rabbits which were immunized with cell-culture adapted FMD virus (A/CHA/09 strain) were used as detector antibody. On the nitrocellulose membrane of the immunochromatographic strip,the capture antibody was laid on a sample pad,the detector antibody was printed at the test line(T) and goat anti-guinea pigs IgG antibodies were immobilized to the control line(C). The lower detection limit of the test for a FMDV 146S antigen is 11.7ng/ml as determined in serial tests after the strip device was assembled and the assay condition optimization. No cross reactions were found with FMDV serotype C,Swine vesicular disease (SVD),Vesicular stomatiti svirus (VSV) and vesicular exanthema of swine virus (VES) viral antigens with this rapid test. Clinically,the diagnostic sensitivity of this test for FMDV serotypes A was 88.7% which is as same as an indirect-sandwich ELISA. The specificity of this strip test was 98.2% and is comparable to the 98.7% obtained with indirect-sandwich ELISA. This rapid strip test is simple,easy and fast for clinical testing on field sites; no special instruments and skills are required,and the result can be obtained within 15 min. To our knowledge,this is the first rapid immunochromatogarpic assay for serotype A of FMDV.
文摘The peptide of amino acids 141～160 of VP1 protein of foot\|and\|mouth disease virus (FMDV) is a major B cell epitope and the peptide of amino acids 21～40 is an important T cell epitope.In this study,the DNA fragments of 141～160 and 21～40 peptide epitopes of a strain of type O FMDV was chemically synthesized and arranged into a tandem repeat 141～160 (20AA)\|21～40 (20AA)\|141～160 (20AA).This tandem sequence was fused to the 3′ end of the heavy chain constant region gene of swine immunoglobulin G and was then cloned into mammalian expression vector pCDM8 to form a recombinant plasmid pCDM8FZ3.After pCDM8FZ3 was inoculated intramuscularly into guinea pigs,it elicited a neutralizing antibody response and a specific spleen T cell proliferative response,and 66% of the vaccinated animals were protected from viral challenge.Our study indicated that the heavy chain constant region of swine IgG can act as the carrier protein for FMDV peptide epitopes,and pCDM8FZ3 is a potential DNA vaccine candidate to prevent FMDV infection.
