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ARS Home » Research » Publications at this Location » Publication #79731
Title: RAPID IDENTIFICATION OF ESCHERICHIA COLI O157:H7 IN BROTH USING A PANEL OF THREE MONOCLONAL ANTIBODIES

Author
item KEEN, JAMES
item HE, YONGSHENG - UNIVERSAL HLTH WATCH INC
item LAEGREID, WILLIAM
item WILSON, RICHARD - PENNSYLVANIA STATE UNIV

Submitted to: Meeting Abstract
Publication Type: Abstract Only
Publication Acceptance Date: 4/21/1997
Publication Date: N/A
Citation: N/A

Interpretive Summary:

Technical Abstract: Monoclonal antibodies (MAbs) generated against lipopolysaccharide (LPS) O-chain and core and against flagella of E. coli O157:H7 were used to accurately serotype bacterial monocultures. Isolates were grown in trypticase soy broth (TSB), heat killed, and directly coated onto replicate wells of microtiter plates. Enzyme-linked immunosorbent assay (ELISA) reactivity following incubation with the anti-O157, anti-LPS core, and anti-H7 MAbs in separate wells permitted independent determination of the presence or absence of the O157, a possible enterohemorrhagic E. coli marker, and the H7 antigens on an isolate. A positive reaction in all 3 wells confirmed the isolate as E. coli O157:H7. The 3 MAb ELISAs had 100% specificity, detected an inoculum of 1 bacterium after 9 h growth at 37øC in TSB, and identified E. coli O157:H7 in a 100-fold excess of non-target bacteria and in artificially contaminated meat. Use of the anti-H7 MAb in an anti-motility assay allowed further verification of H7 expression by E. coli isolates.