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. 1995 Apr;33(4):855–859. doi: 10.1128/jcm.33.4.855-859.1995

Identification of Campylobacter jejuni on the basis of a species-specific gene that encodes a membrane protein.

U Stucki 1, J Frey 1, J Nicolet 1, A P Burnens 1
PMCID: PMC228055  PMID: 7790451

Abstract

To facilitate discrimination between the closely related enteropathogens Campylobacter jejuni and C. coli, unique differences in antigenic surface structure were examined. A genomic library of C. jejuni 81116 was constructed in plasmid pBluescriptIISK- and expressed in Escherichia coli K-12. Rabbit hyperimmune serum raised against C. jejuni ATCC 29428 recognized a clone expressing a C. jejuni 24-kDa membrane-associated protein. Antiserum raised against sonicated recombinant E. coli expressing the 24-kDa protein reacted with C. jejuni, whereas C. coli did not react specifically. Determination of the nucleotide sequence of the DNA insert of this recombinant plasmid revealed an open reading frame encoding 214 amino acids; the gene was designated mapA; and its gene product was designated MAPA. The 18 N-terminal amino acid residues constitute a signal sequence characteristic of prokaryotic membrane lipoproteins. In a dot blot hybridization assay with a mapA probe, 120 clinical isolates of C. jejuni were unequivocally discriminated from 126 other campylobacters, including 34 C. coli isolates. A PCR test based on the mapA sequence was developed for identification of C. jejuni. A PCR product was obtained with all of the clinical isolates of C. jejuni tested from human, dog, cat, bovine calf, and chicken sources. Recombinant MAPA with an added C-terminal six-histidine tail was affinity purified and used to immunize rabbits. The rabbit anti-MAPA serum specifically recognized the protein in whole cells of C. jejuni on Western blots (immunoblots). The MAPA protein was present in all of the C. jejuni strains tested and was absent in C. coli and related campylobacters.

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Selected References

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  1. Blaser M. J., Taylor D. N., Feldman R. A. Epidemiology of Campylobacter jejuni infections. Epidemiol Rev. 1983;5:157–176. doi: 10.1093/oxfordjournals.epirev.a036256. [DOI] [PubMed] [Google Scholar]
  2. Burnens A. P., Nicolet J. Detection of Campylobacter upsaliensis in diarrheic dogs and cats, using a selective medium with cefoperazone. Am J Vet Res. 1992 Jan;53(1):48–51. [PubMed] [Google Scholar]
  3. Camp R. C. Benchmarking--a conversation with Robert C. Camp. The source. Interview by Joe Flower. Healthc Forum J. 1993 Jan-Feb;36(1):30–36. [PubMed] [Google Scholar]
  4. Eyers M., Chapelle S., Van Camp G., Goossens H., De Wachter R. Discrimination among thermophilic Campylobacter species by polymerase chain reaction amplification of 23S rRNA gene fragments. J Clin Microbiol. 1993 Dec;31(12):3340–3343. doi: 10.1128/jcm.31.12.3340-3343.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Giesendorf B. A., Quint W. G., Henkens M. H., Stegeman H., Huf F. A., Niesters H. G. Rapid and sensitive detection of Campylobacter spp. in chicken products by using the polymerase chain reaction. Appl Environ Microbiol. 1992 Dec;58(12):3804–3808. doi: 10.1128/aem.58.12.3804-3808.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Giesendorf B. A., van Belkum A., Koeken A., Stegeman H., Henkens M. H., van der Plas J., Goossens H., Niesters H. G., Quint W. G. Development of species-specific DNA probes for Campylobacter jejuni, Campylobacter coli, and Campylobacter lari by polymerase chain reaction fingerprinting. J Clin Microbiol. 1993 Jun;31(6):1541–1546. doi: 10.1128/jcm.31.6.1541-1546.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Griffiths P. L., Moreno G. S., Park R. W. Differentiation between thermophilic Campylobacter species by species-specific antibodies. J Appl Bacteriol. 1992 Jun;72(6):467–474. doi: 10.1111/j.1365-2672.1992.tb01860.x. [DOI] [PubMed] [Google Scholar]
  8. Hébert G. A., Hollis D. G., Weaver R. E., Lambert M. A., Blaser M. J., Moss C. W. 30 years of campylobacters: biochemical characteristics and a biotyping proposal for Campylobacter jejuni. J Clin Microbiol. 1982 Jun;15(6):1065–1073. doi: 10.1128/jcm.15.6.1065-1073.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Laemmli U. K. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 1970 Aug 15;227(5259):680–685. doi: 10.1038/227680a0. [DOI] [PubMed] [Google Scholar]
  10. March P. E., Inouye M. Characterization of the lep operon of Escherichia coli. Identification of the promoter and the gene upstream of the signal peptidase I gene. J Biol Chem. 1985 Jun 25;260(12):7206–7213. [PubMed] [Google Scholar]
  11. Moreno G. S., Griffiths P. L., Connerton I. F., Park R. W. Occurrence of campylobacters in small domestic and laboratory animals. J Appl Bacteriol. 1993 Jul;75(1):49–54. doi: 10.1111/j.1365-2672.1993.tb03406.x. [DOI] [PubMed] [Google Scholar]
  12. Ng L. K., Stiles M. E., Taylor D. E. Classification of Campylobacter strains using DNA probes. Mol Cell Probes. 1987 Sep;1(3):233–243. doi: 10.1016/0890-8508(87)90036-3. [DOI] [PubMed] [Google Scholar]
  13. Oyofo B. A., Thornton S. A., Burr D. H., Trust T. J., Pavlovskis O. R., Guerry P. Specific detection of Campylobacter jejuni and Campylobacter coli by using polymerase chain reaction. J Clin Microbiol. 1992 Oct;30(10):2613–2619. doi: 10.1128/jcm.30.10.2613-2619.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Penner J. L. The genus Campylobacter: a decade of progress. Clin Microbiol Rev. 1988 Apr;1(2):157–172. doi: 10.1128/cmr.1.2.157. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Pugsley A. P. The complete general secretory pathway in gram-negative bacteria. Microbiol Rev. 1993 Mar;57(1):50–108. doi: 10.1128/mr.57.1.50-108.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Riley M. Functions of the gene products of Escherichia coli. Microbiol Rev. 1993 Dec;57(4):862–952. doi: 10.1128/mr.57.4.862-952.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Salvi R. J., Ahroon W., Saunders S. S., Arnold S. A. Evoked potentials: computer-automated threshold-tracking procedure using an objective detection criterion. Ear Hear. 1987 Jun;8(3):151–156. [PubMed] [Google Scholar]
  18. Sanger F., Nicklen S., Coulson A. R. DNA sequencing with chain-terminating inhibitors. Proc Natl Acad Sci U S A. 1977 Dec;74(12):5463–5467. doi: 10.1073/pnas.74.12.5463. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Tabor S., Richardson C. C. A bacteriophage T7 RNA polymerase/promoter system for controlled exclusive expression of specific genes. Proc Natl Acad Sci U S A. 1985 Feb;82(4):1074–1078. doi: 10.1073/pnas.82.4.1074. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Totten P. A., Patton C. M., Tenover F. C., Barrett T. J., Stamm W. E., Steigerwalt A. G., Lin J. Y., Holmes K. K., Brenner D. J. Prevalence and characterization of hippurate-negative Campylobacter jejuni in King County, Washington. J Clin Microbiol. 1987 Sep;25(9):1747–1752. doi: 10.1128/jcm.25.9.1747-1752.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Vandamme P., Goossens H. Taxonomy of Campylobacter, Arcobacter, and Helicobacter: a review. Zentralbl Bakteriol. 1992 Apr;276(4):447–472. doi: 10.1016/s0934-8840(11)80671-7. [DOI] [PubMed] [Google Scholar]
  22. Vogelstein B., Gillespie D. Preparative and analytical purification of DNA from agarose. Proc Natl Acad Sci U S A. 1979 Feb;76(2):615–619. doi: 10.1073/pnas.76.2.615. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Yamaguchi K., Yu F., Inouye M. A single amino acid determinant of the membrane localization of lipoproteins in E. coli. Cell. 1988 May 6;53(3):423–432. doi: 10.1016/0092-8674(88)90162-6. [DOI] [PubMed] [Google Scholar]

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