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Clinical and Diagnostic Laboratory Immunology, March 1998, p. 242-246, Vol. 5, No. 2
1071-412X/98/$04.00+0
Copyright © 1998, American Society for Microbiology. All rights reserved.

Development of a Blocking Enzyme-Linked Immunosorbent Assay for Detection of Serum Antibodies to O157 Antigen of Escherichia coli

William Laegreid,1,* Mark Hoffman,2 James Keen,1 Robert Elder,1 and Jimmy Kwang1

U.S. Meat Animal Research Center, USDA Agricultural Research Service, Clay Center, Nebraska,1 and National Animal Disease Center, USDA Agricultural Research Service, Ames, Iowa2

Received 11 September 1997/Returned for modification 12 November 1997/Accepted 24 November 1997

The O157 antigen of Escherichia coli shares structural elements with lipopolysaccharide (LPS) antigens of other bacterial species, notably Brucella abortus and Yersinia enterocolitica O9, a fact that confounds the interpretation of assays for anti-O157 antibodies. To address this problem, a blocking enzyme-linked immunosorbent assay (bELISA) was designed with E. coli O157:H7 LPS as the antigen and a monoclonal antibody specific for E. coli O157, designated 13B3, as the competing antibody. The bELISA had equivalent sensitivity to, and significantly higher specificity than, the indirect ELISA (iELISA), detecting anti-O157 antibodies in sera from cattle experimentally inoculated with O157:H7. Only 13% of sera from naive heifers vaccinated for or experimentally infected with B. abortus had increased anti-O157 bELISA titers, while 61% of anti-O157 iELISA titers were increased. The bELISA is a sensitive and specific method for the detection of serum antibodies resulting from exposure to E. coli O157.

* Corresponding author. Mailing address: USDA, ARS, U.S. Meat Animal Research Center, P.O. Box 166, State Spur 18D, Clay Center, NE 68933. Phone: (402) 762-4177. Fax: (402) 762-4375. E-mail: laegreid{at}aux.marc.usda.gov.

Clinical and Diagnostic Laboratory Immunology, March 1998, p. 242-246, Vol. 5, No. 2
1071-412X/98/$04.00+0
Copyright © 1998, American Society for Microbiology. All rights reserved.


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Copyright © 1998 by the American Society for Microbiology. All rights reserved.