Skip to main content
PMC home page
Journal of Clinical Microbiology logoLink to Journal of Clinical Microbiology
. 1993 Jun;31(6):1531–1536. doi: 10.1128/jcm.31.6.1531-1536.1993

Flagellin gene typing of Campylobacter jejuni by restriction fragment length polymorphism analysis.

I Nachamkin 1, K Bohachick 1, C M Patton 1
PMCID: PMC265573  PMID: 8100241

Abstract

We developed and studied a molecular typing approach for Campylobacter spp. with restriction fragment length polymorphism (RFLP) analysis of the flagellin gene flaA in C. jejuni. Using polymerase chain reaction, we amplified the flaA gene from strains comprising different HL:O serotypes by using a primer set directed at the conserved 5' and 3' flaA gene sequence to generate a 1.7-kb amplicon. The amplicon was further digested with the restriction enzyme DdeI, and the fragments generated were analyzed by agarose gel electrophoresis. In 43 non-outbreak strains of six common HL serotypes (HL 1, 2, 4, 5, 9, and 36) in the United States, 18 RFLP patterns were observed. In U.S. outbreak strains previously studied by 10 other typing methods, flaA typing correlated with the HL serotype within each outbreak, and six additional flaA types were identified. Our results suggest that RFLP analysis of the flaA gene from Campylobacter spp. has sufficient discrimination to be useful as a practical typing method for clinical and epidemiologic investigations.

Full text

PDF
1534

Images in this article

1533Image
on p.1533
1535Image
on p.1535
1535Image
on p.1535

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Aeschbacher M., Piffaretti J. C. Population genetics of human and animal enteric Campylobacter strains. Infect Immun. 1989 May;57(5):1432–1437. doi: 10.1128/iai.57.5.1432-1437.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Bolton F. J., Holt A. V., Hutchinson D. N. Campylobacter biotyping scheme of epidemiological value. J Clin Pathol. 1984 Jun;37(6):677–681. doi: 10.1136/jcp.37.6.677. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Bradbury W. C., Marko M. A., Hennessy J. N., Penner J. L. Occurrence of plasmid DNA in serologically defined strains of Campylobacter jejuni and Campylobacter coli. Infect Immun. 1983 May;40(2):460–463. doi: 10.1128/iai.40.2.460-463.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Fischer S. H., Nachamkin I. Common and variable domains of the flagellin gene, flaA, in Campylobacter jejuni. Mol Microbiol. 1991 May;5(5):1151–1158. doi: 10.1111/j.1365-2958.1991.tb01888.x. [DOI] [PubMed] [Google Scholar]
  5. Guerry P., Logan S. M., Thornton S., Trust T. J. Genomic organization and expression of Campylobacter flagellin genes. J Bacteriol. 1990 Apr;172(4):1853–1860. doi: 10.1128/jb.172.4.1853-1860.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Kiehlbauch J. A., Plikaytis B. D., Swaminathan B., Cameron D. N., Wachsmuth I. K. Restriction fragment length polymorphisms in the ribosomal genes for species identification and subtyping of aerotolerant Campylobacter species. J Clin Microbiol. 1991 Aug;29(8):1670–1676. doi: 10.1128/jcm.29.8.1670-1676.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Lior H. New, extended biotyping scheme for Campylobacter jejuni, Campylobacter coli, and "Campylobacter laridis". J Clin Microbiol. 1984 Oct;20(4):636–640. doi: 10.1128/jcm.20.4.636-640.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Lior H., Woodward D. L., Edgar J. A., Laroche L. J., Gill P. Serotyping of Campylobacter jejuni by slide agglutination based on heat-labile antigenic factors. J Clin Microbiol. 1982 May;15(5):761–768. doi: 10.1128/jcm.15.5.761-768.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Nuijten P. J., van Asten F. J., Gaastra W., van der Zeijst B. A. Structural and functional analysis of two Campylobacter jejuni flagellin genes. J Biol Chem. 1990 Oct 15;265(29):17798–17804. [PubMed] [Google Scholar]
  10. Owen R. J., Hernandez J., Bolton F. DNA restriction digest and ribosomal RNA gene patterns of Campylobacter jejuni: a comparison with bio-, sero-, and bacteriophage-types of United Kingdom outbreak strains. Epidemiol Infect. 1990 Oct;105(2):265–275. doi: 10.1017/s0950268800047877. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Patton C. M., Barrett T. J., Morris G. K. Comparison of the Penner and Lior methods for serotyping Campylobacter spp. J Clin Microbiol. 1985 Oct;22(4):558–565. doi: 10.1128/jcm.22.4.558-565.1985. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Patton C. M., Wachsmuth I. K., Evins G. M., Kiehlbauch J. A., Plikaytis B. D., Troup N., Tompkins L., Lior H. Evaluation of 10 methods to distinguish epidemic-associated Campylobacter strains. J Clin Microbiol. 1991 Apr;29(4):680–688. doi: 10.1128/jcm.29.4.680-688.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Penner J. L., Hennessy J. N. Passive hemagglutination technique for serotyping Campylobacter fetus subsp. jejuni on the basis of soluble heat-stable antigens. J Clin Microbiol. 1980 Dec;12(6):732–737. doi: 10.1128/jcm.12.6.732-737.1980. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Salama S. M., Bolton F. J., Hutchinson D. N. Application of a new phagetyping scheme to campylobacters isolated during outbreaks. Epidemiol Infect. 1990 Jun;104(3):405–411. doi: 10.1017/s0950268800047427. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Thornton S. A., Logan S. M., Trust T. J., Guerry P. Polynucleotide sequence relationships among flagellin genes of Campylobacter jejuni and Campylobacter coli. Infect Immun. 1990 Aug;58(8):2686–2689. doi: 10.1128/iai.58.8.2686-2689.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]

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

RESOURCES