• We are sorry, but NCBI web applications do not support your browser and may not function properly. More information
Logo of aemPermissionsJournals.ASM.orgJournalAEM ArticleJournal InfoAuthorsReviewers
Appl Environ Microbiol. Sep 1996; 62(9): 3325–3332.
PMCID: PMC168128

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


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.

Full Text

The Full Text of this article is available as a PDF (482K).

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]

Articles from Applied and Environmental Microbiology are provided here courtesy of American Society for Microbiology (ASM)


Related citations in PubMed

See reviews...See all...

Cited by other articles in PMC

See all...


Recent Activity

Your browsing activity is empty.

Activity recording is turned off.

Turn recording back on

See more...