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KMID : 0357319960310050511
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Journal of the Korean Society for Microbiology
1996 Volume.31 No. 5 p.511 ~ p.524
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Production of Monoclonal Antibodies Specific for Salmonella typhi Vi Antigen and Their Characterization and Application of Rapid Detection Method
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이영희
주영란/김혜진/정하용/박강수/ㄱ박기덕/황규잠/이광준/윤희주
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Abstract
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Serum antibodies to Vi antigen were detected in mice immunized with the purified Vi antigen from Salmonella typhi whole cells. Fusion of the spleen cells from one of the Vi antibody producing mice with myeloma cells produced two stable hybridomas
that
secreted antibodies to Vi. Produced two monoclonal antibodies(SM-Vi1, SM-Vi2) were all of the immunoglobulin M class. As determined by ELISA, slide agglutination test, and immunoblotting, they appeared to have the specificity to Vi antigen Mouse
ascitic
fluids induced by monoclonal antibody(SM-Vi1) were used in a direct slide agglutination test. SM-Vi1 could sharply differentiate the S. typhi from various Salmonella serotypes(129 strains). And it also showed no cross-reactivity to other enteric
bacteria(105 strains belonging to 5 genus) which were isolated from clinical specimens. Therefore. these results suggest that the SM-Vi1 monoclonal antibody could be developed as S. typhi Vi diagnostic antisera. We obtained 5.4mg of purified
monoclonal
antibody(SM-Vi1) from mice ascitic fluid 6ml and its minimal detectable concentration was 99ng/ml in 10E6 cell/ml by ELISA. Using the modified double antibody sandwich ELISA, purified monoclonal antibody SM-Vi1 was applied to detect S. typhi and
E.
coli
in artificially inoculated(5×10E6cells/ml) subterranean water specimen. Purified monoclonal antibody was reacted only to against S. typhi but not to against E. coli. Thus, SM-Vi1 monoclonal antibody, Using the modified double antibody sandwich
ELISA
seems to be useful method in detecting S. typhi in environment water specimen.
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