Logo of jcmPermissionsJournals.ASM.orgJournalJCM ArticleJournal InfoAuthorsReviewers
J Clin Microbiol. 1992 May; 30(5): 1189–1193.
PMCID: PMC265248

Rapid and specific detection of the pap, afa, and sfa adhesin-encoding operons in uropathogenic Escherichia coli strains by polymerase chain reaction.


Adhesin-encoding operons (pap, sfa/foc, and afa) have been shown to be prevalent in Escherichia coli strains associated with urinary tract infections. A quick and sensitive assay to identify these operons was developed by using the polymerase chain reaction (PCR). Three pairs of 25-mer primers were defined from the sequences of the DNA fragments used as probes in hybridization studies to identify each of the three operons, and the six primers were used together in a single reaction of amplification. To validate the PCR approach for detection of adhesin-encoding operons among clinical isolates, we investigated a collection of 97 E. coli isolates with the following characteristics: all isolates originated from the urine of patients with pyelonephritis, and the adhesin responsible for specific binding of the isolates to uroepithelial cells was previously characterized by phenotypic assays, as well as genotypic tests based on hybridization. There was a perfect correlation between the results obtained with the PCR approach and those previously obtained by using DNA probes. These results indicate that the PCR method, which is highly specific and easier to perform than the hybridization method, is a powerful genotypic assay for detection of adhesin-encoding operons. Thus, this assay can be recommended for clinical use to detect virulent urinary E. coli strains, as well as for epidemiological studies.

Full text

Full text is available as a scanned copy of the original print version. Get a printable copy (PDF file) of the complete article (1.0M), or click on a page image below to browse page by page. Links to PubMed are also available for Selected References.

Images in this article

Click on the image to see a larger version.

Selected References

These references are in PubMed. This may not be the complete list of references from this article.
  • Archambaud M, Courcoux P, Labigne-Roussel A. Detection by molecular hybridization of pap, afa, and sfa adherence systems in Escherichia coli strains associated with urinary and enteral infections. Ann Inst Pasteur Microbiol. 1988 Sep-Oct;139(5):575–588. [PubMed]
  • Archambaud M, Courcoux P, Ouin V, Chabanon G, Labigne-Roussel A. Phenotypic and genotypic assays for the detection and identification of adhesins from pyelonephritic Escherichia coli. Ann Inst Pasteur Microbiol. 1988 Sep-Oct;139(5):557–573. [PubMed]
  • Bilge SS, Clausen CR, Lau W, Moseley SL. Molecular characterization of a fimbrial adhesin, F1845, mediating diffuse adherence of diarrhea-associated Escherichia coli to HEp-2 cells. J Bacteriol. 1989 Aug;171(8):4281–4289. [PMC free article] [PubMed]
  • Bolivar F, Rodriguez RL, Greene PJ, Betlach MC, Heyneker HL, Boyer HW, Crosa JH, Falkow S. Construction and characterization of new cloning vehicles. II. A multipurpose cloning system. Gene. 1977;2(2):95–113. [PubMed]
  • Boyer HW, Roulland-Dussoix D. A complementation analysis of the restriction and modification of DNA in Escherichia coli. J Mol Biol. 1969 May 14;41(3):459–472. [PubMed]
  • Buchanan K, Falkow S, Hull RA, Hull SI. Frequency among Enterobacteriaceae of the DNA sequences encoding type 1 pili. J Bacteriol. 1985 May;162(2):799–803. [PMC free article] [PubMed]
  • Dowling KJ, Roberts JA, Kaack MB. P-fimbriated Escherichia coli urinary tract infection: a clinical correlation. South Med J. 1987 Dec;80(12):1533–1536. [PubMed]
  • Fowler JE, Jr, Stamey TA. Studies of introital colonization in women with recurrent urinary infections. VII. The role of bacterial adherence. J Urol. 1977 Apr;117(4):472–476. [PubMed]
  • Frankel G, Giron JA, Valmassoi J, Schoolnik GK. Multi-gene amplification: simultaneous detection of three virulence genes in diarrhoeal stool. Mol Microbiol. 1989 Dec;3(12):1729–1734. [PubMed]
  • Gander RM, Thomas VL, Forland M. Mannose-resistant hemagglutination and P receptor recognition of uropathogenic Escherichia coli isolated from adult patients. J Infect Dis. 1985 Mar;151(3):508–513. [PubMed]
  • Göransson M, Uhlin BE. Environmental temperature regulates transcription of a virulence pili operon in E. coli. EMBO J. 1984 Dec 1;3(12):2885–2888. [PMC free article] [PubMed]
  • Grüneberg RN. Antibiotic sensitivities of urinary pathogens, 1971-82. J Antimicrob Chemother. 1984 Jul;14(1):17–23. [PubMed]
  • Hacker J, Schmidt G, Hughes C, Knapp S, Marget M, Goebel W. Cloning and characterization of genes involved in production of mannose-resistant, neuraminidase-susceptible (X) fimbriae from a uropathogenic O6:K15:H31 Escherichia coli strain. Infect Immun. 1985 Feb;47(2):434–440. [PMC free article] [PubMed]
  • Hull RA, Gill RE, Hsu P, Minshew BH, Falkow S. Construction and expression of recombinant plasmids encoding type 1 or D-mannose-resistant pili from a urinary tract infection Escherichia coli isolate. Infect Immun. 1981 Sep;33(3):933–938. [PMC free article] [PubMed]
  • Johnson JR. Virulence factors in Escherichia coli urinary tract infection. Clin Microbiol Rev. 1991 Jan;4(1):80–128. [PMC free article] [PubMed]
  • Johnson JR, Stamm WE. Urinary tract infections in women: diagnosis and treatment. Ann Intern Med. 1989 Dec 1;111(11):906–917. [PubMed]
  • Källenius G, Möllby R, Svenson SB, Helin I, Hultberg H, Cedergren B, Winberg J. Occurrence of P-fimbriated Escherichia coli in urinary tract infections. Lancet. 1981 Dec 19;2(8260-61):1369–1372. [PubMed]
  • Labigne-Roussel A, Falkow S. Distribution and degree of heterogeneity of the afimbrial-adhesin-encoding operon (afa) among uropathogenic Escherichia coli isolates. Infect Immun. 1988 Mar;56(3):640–648. [PMC free article] [PubMed]
  • Labigne-Roussel AF, Lark D, Schoolnik G, Falkow S. Cloning and expression of an afimbrial adhesin (AFA-I) responsible for P blood group-independent, mannose-resistant hemagglutination from a pyelonephritic Escherichia coli strain. Infect Immun. 1984 Oct;46(1):251–259. [PMC free article] [PubMed]
  • Lomberg H, Hellström M, Jodal U, Leffler H, Lincoln K, Svanborg Edén C. Virulence-associated traits in Escherichia coli causing first and recurrent episodes of urinary tract infection in children with or without vesicoureteral reflux. J Infect Dis. 1984 Oct;150(4):561–569. [PubMed]
  • Low D, Robinson EN, Jr, McGee ZA, Falkow S. The frequency of expression of pyelonephritis-associated pili is under regulatory control. Mol Microbiol. 1987 Nov;1(3):335–346. [PubMed]
  • Lund B, Marklund BI, Strömberg N, Lindberg F, Karlsson KA, Normark S. Uropathogenic Escherichia coli can express serologically identical pili of different receptor binding specificities. Mol Microbiol. 1988 Mar;2(2):255–263. [PubMed]
  • Meyrier A, Condamin MC, Fernet M, Labigne-Roussel A, Simon P, Callard P, Rainfray M, Soilleux M, Groc A. Frequency of development of early cortical scarring in acute primary pyelonephritis. Kidney Int. 1989 Feb;35(2):696–703. [PubMed]
  • Norgren M, Båga M, Tennent JM, Normark S. Nucleotide sequence, regulation and functional analysis of the papC gene required for cell surface localization of Pap pili of uropathogenic Escherichia coli. Mol Microbiol. 1987 Sep;1(2):169–178. [PubMed]
  • Norgren M, Normark S, Lark D, O'Hanley P, Schoolnik G, Falkow S, Svanborg-Edén C, Båga M, Uhlin BE. Mutations in E coli cistrons affecting adhesion to human cells do not abolish Pap pili fiber formation. EMBO J. 1984 May;3(5):1159–1165. [PMC free article] [PubMed]
  • O'Hanley P, Lark D, Falkow S, Schoolnik G. Molecular basis of Escherichia coli colonization of the upper urinary tract in BALB/c mice. Gal-Gal pili immunization prevents Escherichia coli pyelonephritis in the BALB/c mouse model of human pyelonephritis. J Clin Invest. 1985 Feb;75(2):347–360. [PMC free article] [PubMed]
  • O'Hanley P, Low D, Romero I, Lark D, Vosti K, Falkow S, Schoolnik G. Gal-Gal binding and hemolysin phenotypes and genotypes associated with uropathogenic Escherichia coli. N Engl J Med. 1985 Aug 15;313(7):414–420. [PubMed]
  • Ott M, Schmoll T, Goebel W, Van Die I, Hacker J. Comparison of the genetic determinant coding for the S-fimbrial adhesin (sfa) of Escherichia coli to other chromosomally encoded fimbrial determinants. Infect Immun. 1987 Aug;55(8):1940–1943. [PMC free article] [PubMed]
  • Pere A, Nowicki B, Saxén H, Siitonen A, Korhonen TK. Expression of P, type-1, and type-1C fimbriae of Escherichia coli in the urine of patients with acute urinary tract infection. J Infect Dis. 1987 Oct;156(4):567–574. [PubMed]
  • Rhen M, Väisänen-Rhen V, Saraste M, Korhonen TK. Organization of genes expressing the blood-group-M-specific hemagglutinin of Escherichia coli: identification and nucleotide sequence of the M-agglutinin subunit gene. Gene. 1986;49(3):351–360. [PubMed]
  • Riegman N, Kusters R, Van Veggel H, Bergmans H, Van Bergen en Henegouwen P, Hacker J, Van Die I. F1C fimbriae of a uropathogenic Escherichia coli strain: genetic and functional organization of the foc gene cluster and identification of minor subunits. J Bacteriol. 1990 Feb;172(2):1114–1120. [PMC free article] [PubMed]
  • Sanger F, Coulson AR, Barrell BG, Smith AJ, Roe BA. Cloning in single-stranded bacteriophage as an aid to rapid DNA sequencing. J Mol Biol. 1980 Oct 25;143(2):161–178. [PubMed]
  • Sanger F, Nicklen S, Coulson AR. DNA sequencing with chain-terminating inhibitors. Proc Natl Acad Sci U S A. 1977 Dec;74(12):5463–5467. [PMC free article] [PubMed]
  • Schmoll T, Morschhäuser J, Ott M, Ludwig B, van Die I, Hacker J. Complete genetic organization and functional aspects of the Escherichia coli S fimbrial adhesion determinant: nucleotide sequence of the genes sfa B, C, D, E, F. Microb Pathog. 1990 Nov;9(5):331–343. [PubMed]
  • Edén CS, Hanson LA, Jodal U, Lindberg U, Akerlund AS. Variable adherence to normal human urinary-tract epithelial cells of Escherichia coli strains associated with various forms of urinary-tract infection. Lancet. 1976 Sep 4;1(7984):490–492. [PubMed]
  • Swanson TN, Bilge SS, Nowicki B, Moseley SL. Molecular structure of the Dr adhesin: nucleotide sequence and mapping of receptor-binding domain by use of fusion constructs. Infect Immun. 1991 Jan;59(1):261–268. [PMC free article] [PubMed]
  • Väisänen-Rhen V, Elo J, Väisänen E, Siitonen A, Orskov I, Orskov F, Svenson SB, Mäkelä PH, Korhonen TK. P-fimbriated clones among uropathogenic Escherichia coli strains. Infect Immun. 1984 Jan;43(1):149–155. [PMC free article] [PubMed]
  • Väisänen V, Elo J, Tallgren LG, Siitonen A, Mäkelä PH, Svanborg-Edén C, Källenius G, Svenson SB, Hultberg H, Korhonen T. Mannose-resistant haemagglutination and P antigen recognition are characteristic of Escherichia coli causing primary pyelonephritis. Lancet. 1981 Dec 19;2(8260-61):1366–1369. [PubMed]
  • Westerlund B, Kuusela P, Risteli J, Risteli L, Vartio T, Rauvala H, Virkola R, Korhonen TK. The O75X adhesin of uropathogenic Escherichia coli is a type IV collagen-binding protein. Mol Microbiol. 1989 Mar;3(3):329–337. [PubMed]
  • Yanisch-Perron C, Vieira J, Messing J. Improved M13 phage cloning vectors and host strains: nucleotide sequences of the M13mp18 and pUC19 vectors. Gene. 1985;33(1):103–119. [PubMed]

Articles from Journal of Clinical Microbiology are provided here courtesy of American Society for Microbiology (ASM)


Save items

Related citations in PubMed

See reviews...See all...

Cited by other articles in PMC

See all...


  • MedGen
    Related information in MedGen
  • PubMed
    PubMed citations for these articles

Recent Activity

Your browsing activity is empty.

Activity recording is turned off.

Turn recording back on

See more...