• We are sorry, but NCBI web applications do not support your browser and may not function properly. More information
Logo of jcmPermissionsJournals.ASM.orgJournalJCM ArticleJournal InfoAuthorsReviewers
J Clin Microbiol. Oct 1997; 35(10): 2568–2572.
PMCID: PMC230012

PCR detection, identification to species level, and fingerprinting of Campylobacter jejuni and Campylobacter coli direct from diarrheic samples.

Abstract

Three sets of primers were designed for PCR detection and differentiation of Campylobacter jejuni and Campylobacter coli. The first PCR assay was designed to coidentify C. jejuni and C. coli based on their 16S rRNA gene sequences. The second PCR assay, based on the hippuricase gene sequence, identified all tested reference strains of C. jejuni and also strains of that species which lack detectable hippuricase activity. The third PCR assay, based on the sequence of a cloned (putative) aspartokinase gene and the downstream open reading frame, identified all tested reference strains of C. coli. The assays will find immediate application in the rapid identification to species level of isolates. The assays combine with a protocol for purification of total DNA from fecal samples to allow reproducible PCR identification of campylobacters directly from stools. Of 20 clinical samples from which campylobacters had been cultured, we detected C. jejuni in 17, C. coli in 2, and coinfection of C. jejuni and Campylobacter hyointestinalis in 1. These results were concordant with culture and phenotypic identification to species level. Strain typing by PCR-restriction fragment length polymorphism of the flagellin (flaA) gene detected identical flaA types in fecal DNA and the corresponding campylobacter isolate. Twenty-five Campylobacter-negative stool samples gave no reaction with the PCR assays. These PCR assays can rapidly define the occurrence, species incidence, and flaA genotypes of enteropathogenic campylobacters.

Full Text

The Full Text of this article is available as a PDF (596K).

Selected References

These references are in PubMed. This may not be the complete list of references from this article.
  • Alderton MR, Korolik V, Coloe PJ, Dewhirst FE, Paster BJ. Campylobacter hyoilei sp. nov., associated with porcine proliferative enteritis. Int J Syst Bacteriol. 1995 Jan;45(1):61–66. [PubMed]
  • Alm RA, Guerry P, Trust TJ. Distribution and polymorphism of the flagellin genes from isolates of Campylobacter coli and Campylobacter jejuni. J Bacteriol. 1993 May;175(10):3051–3057. [PMC free article] [PubMed]
  • Berg RD. The indigenous gastrointestinal microflora. Trends Microbiol. 1996 Nov;4(11):430–435. [PubMed]
  • Boom R, Sol CJ, Salimans MM, Jansen CL, Wertheim-van Dillen PM, van der Noordaa J. Rapid and simple method for purification of nucleic acids. J Clin Microbiol. 1990 Mar;28(3):495–503. [PMC free article] [PubMed]
  • Edmonds P, Patton CM, Griffin PM, Barrett TJ, Schmid GP, Baker CN, Lambert MA, Brenner DJ. Campylobacter hyointestinalis associated with human gastrointestinal disease in the United States. J Clin Microbiol. 1987 Apr;25(4):685–691. [PMC free article] [PubMed]
  • 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. [PMC free article] [PubMed]
  • Fennell CL, Rompalo AM, Totten PA, Bruch KL, Flores BM, Stamm WE. Isolation of "Campylobacter hyointestinalis" from a human. J Clin Microbiol. 1986 Jul;24(1):146–148. [PMC free article] [PubMed]
  • Giesendorf BA, Quint WG, Henkens MH, Stegeman H, Huf FA, Niesters HG. 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. [PMC free article] [PubMed]
  • Hani EK, Chan VL. Expression and characterization of Campylobacter jejuni benzoylglycine amidohydrolase (Hippuricase) gene in Escherichia coli. J Bacteriol. 1995 May;177(9):2396–2402. [PMC free article] [PubMed]
  • Kendall EJ, Tanner EI. Campylobacter enteritis in general practice. J Hyg (Lond) 1982 Apr;88(2):155–163. [PMC free article] [PubMed]
  • Linton D, Dewhirst FE, Clewley JP, Owen RJ, Burnens AP, Stanley J. Two types of 16S rRNA gene are found in Campylobacter helveticus: analysis, applications and characterization of the intervening sequence found in some strains. Microbiology. 1994 Apr;140(Pt 4):847–855. [PubMed]
  • Linton D, Owen RJ, Stanley J. Rapid identification by PCR of the genus Campylobacter and of five Campylobacter species enteropathogenic for man and animals. Res Microbiol. 1996 Nov-Dec;147(9):707–718. [PubMed]
  • Minet J, Grosbois B, Megraud F. Campylobacter hyointestinalis: an opportunistic enteropathogen? J Clin Microbiol. 1988 Dec;26(12):2659–2660. [PMC free article] [PubMed]
  • Nicholson MA, Patton CM. Application of Lior biotyping by use of genetically identified Campylobacter strains. J Clin Microbiol. 1993 Dec;31(12):3348–3350. [PMC free article] [PubMed]
  • Oyofo BA, Thornton SA, Burr DH, Trust TJ, Pavlovskis OR, Guerry P. Specific detection of Campylobacter jejuni and Campylobacter coli by using polymerase chain reaction. J Clin Microbiol. 1992 Oct;30(10):2613–2619. [PMC free article] [PubMed]
  • Roop RM, 2nd, Smibert RM, Johnson JL, Krieg NR. Differential characteristics of catalase-positive campylobacters correlated with DNA homology groups. Can J Microbiol. 1984 Jul;30(7):938–951. [PubMed]
  • Stanley J, Linton D, Sutherland K, Jones C, Owen RJ. High-resolution genotyping of Campylobacter coli identifies clones of epidemiologic and evolutionary significance. J Infect Dis. 1995 Oct;172(4):1130–1134. [PubMed]
  • Stucki U, Frey J, Nicolet J, Burnens AP. Identification of Campylobacter jejuni on the basis of a species-specific gene that encodes a membrane protein. J Clin Microbiol. 1995 Apr;33(4):855–859. [PMC free article] [PubMed]
  • Camp RC. Benchmarking--a conversation with Robert C. Camp. The source. Interview by Joe Flower. Healthc Forum J. 1993 Jan-Feb;36(1):30–36. [PubMed]
  • Waegel A, Nachamkin I. Detection and molecular typing of Campylobacter jejuni in fecal samples by polymerase chain reaction. Mol Cell Probes. 1996 Apr;10(2):75–80. [PubMed]
  • Wegmüller B, Lüthy J, Candrian U. Direct polymerase chain reaction detection of Campylobacter jejuni and Campylobacter coli in raw milk and dairy products. Appl Environ Microbiol. 1993 Jul;59(7):2161–2165. [PMC free article] [PubMed]
  • Widjojoatmodjo MN, Fluit AC, Torensma R, Verdonk GP, Verhoef J. The magnetic immuno polymerase chain reaction assay for direct detection of salmonellae in fecal samples. J Clin Microbiol. 1992 Dec;30(12):3195–3199. [PMC free article] [PubMed]

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

Formats:

Related citations in PubMed

See reviews...See all...

Cited by other articles in PMC

See all...

Links

Recent Activity

Your browsing activity is empty.

Activity recording is turned off.

Turn recording back on

See more...