Author
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RIVERA BETANCOURT, MILDRED |
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KEEN, JAMES |
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YANG, SIMON - FORMER ARS EMPLOYEE |
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Submitted to: American Society for Microbiology
Publication Type: Abstract Only Publication Acceptance Date: 2/25/1998 Publication Date: N/A Citation: N/A Interpretive Summary: Technical Abstract: Escherichia coli is the most frequently isolated bacteria from all types of urinary tract infections. Uropathogenic strains exhibit a limited number of lipopolysaccharide (LPS O), capsular acidic polysaccharide (K), and flagellar protein (H) antigens which often codistribute with other virulence factors. E. coli O4:H5 is one of the most common uropathogenic serotypes. In order to facilitate identification and detection of uropathogenic E. coli, two monoclonal antibodies (MAbs 2C5-F10 and 8D1) of the IgM and IgG1 isotypes, respectively, were produced from splenocytes of BALB/c mice injected with E. coli O4:K3:H5 partially-purified flagella. A total of 119 E. coli strains, 48 Salmonella spp. serovars, and 25 other Gram-negative bacteria were used to evaluate the specificity of the MAbs by ELISA. The MAb 2C5-F10 reacted specifically with E. coli serotype O4-H5 and not with the other non-O4:H5 tested. MAb 8D1 reacted with 4 of 5 E. coli H5 isolates and also cross-reacted strongly with H56 serotype on ELISA. SDS-PAGE of whole cell lysates of E. coli O4:H5, H56, and O4:NM serotypes followed by Western immunoblotting revealed that MAb 8D1 reacted with all the E. coli H5 strains but not with H56 serotype. Western blot analysis using MAb 2C5-F10 further supported specificity of this MAb for the O4:H5 serotype and showed a ladder pattern consistent with an LPS-associated epitope. The apparent specificity of MAb 2C5-F10 may make it a useful immunoreagent for the detection of O4:H5 uropathogens in clinical specimens while MAb 8D1 could be a useful tool for H5 serotyping. |
