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Appl Environ Microbiol. Sep 1996; 62(9): 3325–3332.
PMCID: PMC168128

Monoclonal antibodies for detection of the H7 antigen of Escherichia coli.

Abstract

Two murine monoclonal antibodies (MAbs) (2B7 and 46E9-9) reactive with the H7 flagellar antigen of Escherichia coli were produced and characterized. A total of 217 E. coli strains (48 O157:H7, 4 O157:NM, 23 O157:non-H7, 22 H7:non-O157, and 120 non-O157:nonH7), 17 Salmonella serovars, and 29 other gram-negative bacteria were used to evaluate the reactivities of the two MAbs by indirect enzyme-linked immunosorbent assay (ELISA). Both MAbs reacted strongly with all E. coli strains possessing the H7 antigen and with H23- and H24-positive E. coli strains. Indirect ELISA MAb specificity was confirmed by inhibition ELISA and by Western blotting (immunoblotting), using partially purified flagellins from E. coli O157:H7 and other E. coli strains. On a Western blot, MAb 46E9-9 was more reactive against H7 flagellin of E. coli O157:H7 than against H7 flagellin of E. coli O1:K1:H7. Competition ELISA suggested that MAbs 2B7 and 46E9-9 reacted with closely related H7 epitopes. When the ELISA reactivities of the MAbs and two commercially available polyclonal anti-H7 antisera were compared, both polyclonal antisera and MAbs reacted strongly with E. coli H7 bacteria. However, the polyclonal antisera cross-reacted strongly both with non-H7 E. coli and with many non-E. coli bacteria. The polyclonal antisera also reacted strongly with H23 and H24 E. coli isolates. The data suggest the need to define serotype-specific epitopes among H7, H23, and H24 E. coli flagella. The anti-H7 MAbs described in this report have the potential to serve as high-quality diagnostic reagents, used either alone or in combination with O157-specific MAbs, to identify or detect E. coli O157:H7 in food products or in human and veterinary clinical specimens.

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Selected References

These references are in PubMed. This may not be the complete list of references from this article.
  • Carter AO, Borczyk AA, Carlson JA, Harvey B, Hockin JC, Karmali MA, Krishnan C, Korn DA, Lior H. A severe outbreak of Escherichia coli O157:H7--associated hemorrhagic colitis in a nursing home. N Engl J Med. 1987 Dec 10;317(24):1496–1500. [PubMed]
  • Erich T, Schellekens J, Bouter A, Van Kranen J, Brouwer E, Verhoef J. Binding characteristics and cross-reactivity of three different antilipid A monoclonal antibodies. J Immunol. 1989 Dec 15;143(12):4053–4060. [PubMed]
  • Evins GM, Gheesling LL, Tauxe RV. Quality of commercially produced Shigella serogrouping and serotyping antisera. J Clin Microbiol. 1988 Mar;26(3):438–442. [PMC free article] [PubMed]
  • Farmer JJ, 3rd, Davis BR. H7 antiserum-sorbitol fermentation medium: a single tube screening medium for detecting Escherichia coli O157:H7 associated with hemorrhagic colitis. J Clin Microbiol. 1985 Oct;22(4):620–625. [PMC free article] [PubMed]
  • Fujita H, Yamaguchi S, Taira T, Iino T. A simple method for the isolation of flagellar shape mutants in Salmonella. J Gen Microbiol. 1981 Jul;125(1):213–216. [PubMed]
  • Gransden WR, Damm MA, Anderson JD, Carter JE, Lior H. Further evidence associating hemolytic uremic syndrome with infection by Verotoxin-producing Escherichia coli O157:H7. J Infect Dis. 1986 Sep;154(3):522–524. [PubMed]
  • Griffin PM, Tauxe RV. The epidemiology of infections caused by Escherichia coli O157:H7, other enterohemorrhagic E. coli, and the associated hemolytic uremic syndrome. Epidemiol Rev. 1991;13:60–98. [PubMed]
  • Hancock DD, Besser TE, Kinsel ML, Tarr PI, Rice DH, Paros MG. The prevalence of Escherichia coli O157.H7 in dairy and beef cattle in Washington State. Epidemiol Infect. 1994 Oct;113(2):199–207. [PMC free article] [PubMed]
  • He Y, Keel BA. Biochemical characterization of bovine alpha-fetoprotein and comparison with human alpha-fetoprotein. Comp Biochem Physiol Biochem Mol Biol. 1994 Jul;108(3):327–336. [PubMed]
  • Johnson RP, Durham RJ, Johnson ST, MacDonald LA, Jeffrey SR, Butman BT. Detection of Escherichia coli O157:H7 in meat by an enzyme-linked immunosorbent assay, EHEC-Tek. Appl Environ Microbiol. 1995 Jan;61(1):386–388. [PMC free article] [PubMed]
  • Karmali MA. Infection by verocytotoxin-producing Escherichia coli. Clin Microbiol Rev. 1989 Jan;2(1):15–38. [PMC free article] [PubMed]
  • Karmali MA, Petric M, Lim C, Fleming PC, Steele BT. Escherichia coli cytotoxin, haemolytic-uraemic syndrome, and haemorrhagic colitis. Lancet. 1983 Dec 3;2(8362):1299–1300. [PubMed]
  • Karmali MA, Steele BT, Petric M, Lim C. Sporadic cases of haemolytic-uraemic syndrome associated with faecal cytotoxin and cytotoxin-producing Escherichia coli in stools. Lancet. 1983 Mar 19;1(8325):619–620. [PubMed]
  • Kim MS, Doyle MP. Dipstick immunoassay to detect enterohemorrhagic Escherichia coli O157:H7 in retail ground beef. Appl Environ Microbiol. 1992 May;58(5):1764–1767. [PMC free article] [PubMed]
  • Lawn AM, Orskov I, Orskov F. Morphological distinction between different H serotypes of Escherichia coli. J Gen Microbiol. 1977 Jul;101(1):111–119. [PubMed]
  • MacDonald KL, O'Leary MJ, Cohen ML, Norris P, Wells JG, Noll E, Kobayashi JM, Blake PA. Escherichia coli O157:H7, an emerging gastrointestinal pathogen. Results of a one-year, prospective, population-based study. JAMA. 1988 Jun 24;259(24):3567–3570. [PubMed]
  • Mills KW, Tietze KL. Monoclonal antibody enzyme-linked immunosorbent assay for identification of K99-positive Escherichia coli isolates from calves. J Clin Microbiol. 1984 Apr;19(4):498–501. [PMC free article] [PubMed]
  • Padhye NV, Doyle MP. Rapid procedure for detecting enterohemorrhagic Escherichia coli O157:H7 in food. Appl Environ Microbiol. 1991 Sep;57(9):2693–2698. [PMC free article] [PubMed]
  • Padhye NV, Doyle MP. Production and characterization of a monoclonal antibody specific for enterohemorrhagic Escherichia coli of serotypes O157:H7 and O26:H11. J Clin Microbiol. 1991 Jan;29(1):99–103. [PMC free article] [PubMed]
  • Padhye VV, Zhao T, Doyle MP. Production and characterisation of monoclonal antibodies to Verotoxins 1 and 2 from Escherichia coli of serotype O 157:H7. J Med Microbiol. 1989 Nov;30(3):219–226. [PubMed]
  • Perera LP, Marques LR, O'Brien AD. Isolation and characterization of monoclonal antibodies to Shiga-like toxin II of enterohemorrhagic Escherichia coli and use of the monoclonal antibodies in a colony enzyme-linked immunosorbent assay. J Clin Microbiol. 1988 Oct;26(10):2127–2131. [PMC free article] [PubMed]
  • Perry MB, Bundle DR, Gidney MA, Lior H. Identification of Escherichia coli serotype O157 strains by using a monoclonal antibody. J Clin Microbiol. 1988 Nov;26(11):2391–2394. [PMC free article] [PubMed]
  • Remis RS, MacDonald KL, Riley LW, Puhr ND, Wells JG, Davis BR, Blake PA, Cohen ML. Sporadic cases of hemorrhagic colitis associated with Escherichia coli O157:H7. Ann Intern Med. 1984 Nov;101(5):624–626. [PubMed]
  • Riley LW, Remis RS, Helgerson SD, McGee HB, Wells JG, Davis BR, Hebert RJ, Olcott ES, Johnson LM, Hargrett NT, et al. Hemorrhagic colitis associated with a rare Escherichia coli serotype. N Engl J Med. 1983 Mar 24;308(12):681–685. [PubMed]
  • Spika JS, Parsons JE, Nordenberg D, Wells JG, Gunn RA, Blake PA. Hemolytic uremic syndrome and diarrhea associated with Escherichia coli O157:H7 in a day care center. J Pediatr. 1986 Aug;109(2):287–291. [PubMed]
  • Strockbine NA, Marques LR, Holmes RK, O'Brien AD. Characterization of monoclonal antibodies against Shiga-like toxin from Escherichia coli. Infect Immun. 1985 Dec;50(3):695–700. [PMC free article] [PubMed]
  • Szabo R, Todd E, MacKenzie J, Parrington L, Armstrong A. Increased sensitivity of the rapid hydrophobic grid membrane filter enzyme-labeled antibody procedure for Escherichia coli O157 detection in foods and bovine feces. Appl Environ Microbiol. 1990 Nov;56(11):3546–3549. [PMC free article] [PubMed]
  • Tison DL. Culture confirmation of Escherichia coli serotype O157:H7 by direct immunofluorescence. J Clin Microbiol. 1990 Mar;28(3):612–613. [PMC free article] [PubMed]
  • Willshaw GA, Thirlwell J, Jones AP, Parry S, Salmon RL, Hickey M. Vero cytotoxin-producing Escherichia coli O157 in beefburgers linked to an outbreak of diarrhoea, haemorrhagic colitis and haemolytic uraemic syndrome in Britain. Lett Appl Microbiol. 1994 Nov;19(5):304–307. [PubMed]

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