Development of a m‐PCR assay for simultaneous identification of Campylobacter jejuni and C. coli

Multiplex PCR assay (m–PCR) with three sets of primers was developed for simultaneous identification of Campylobacter jejuni and C. coli. Poultry faecal samples were enriched in Preston broth for 24 h and streaking on selective media was performed before and after enrichment. m–PCR was applied on bacterial cultures harvested from media plates. The data showed a selective effect of Preston broth which favoured the growth of C. coli. Identification of the species by the hippurate hydrolysis test and by the m–PCR was performed on 294 isolates of Campylobacter. The efficiency of the identification by the biochemical test is only 34% in comparison to 100% efficiency with the PCR. The use of our m–PCR in combination with the culture method allowed reliable detection and identification of C. jejuni and C. coli within 3–4 d.

[1]  J. Stanley,et al.  Detection of campylobacter in gastroenteritis: comparison of direct PCR assay of faecal samples with selective culture , 1998, Epidemiology and Infection.

[2]  J. Corry,et al.  Evaluation of a new arcobacter enrichment medium and comparison with two media developed for enrichment of Campylobacter spp. , 1998, International journal of food microbiology.

[3]  D. Linton,et al.  PCR detection, identification to species level, and fingerprinting of Campylobacter jejuni and Campylobacter coli direct from diarrheic samples , 1997, Journal of clinical microbiology.

[4]  Thomas L. Madden,et al.  Gapped BLAST and PSI-BLAST: a new generation of protein database search programs. , 1997, Nucleic acids research.

[5]  R. Auckenthaler,et al.  Etiological agents of infectious diarrhea: implications for requests for microbial culture , 1997, Journal of clinical microbiology.

[6]  K. Harmon,et al.  Differentiation of Campylobacter jejuni and Campylobacter coli by polymerase chain reaction. , 1997, Molecular and cellular probes.

[7]  M. Collins,et al.  Specific identification of the enteropathogens Campylobacter jejuni and Campylobacter coli by using a PCR test based on the ceuE gene encoding a putative virulence determinant , 1997, Journal of clinical microbiology.

[8]  L. Sicinschi [The comparative identification of Campylobacter strains by traditional enzymatic tests and the gene amplification reaction]. , 1995, Bacteriologia, virusologia, parazitologia, epidemiologia.

[9]  J. Nicolet,et al.  Identification of Campylobacter jejuni on the basis of a species-specific gene that encodes a membrane protein , 1995, Journal of clinical microbiology.

[10]  C. Patton,et al.  Application of Lior biotyping by use of genetically identified Campylobacter strains , 1993, Journal of clinical microbiology.

[11]  H. Goossens,et al.  Campylobacter and helicobacter infections , 1992 .

[12]  G. Morris,et al.  Differentiation of Campylobacter Species Using Phenotypic Characterization , 1988 .

[13]  K. Holmes,et al.  Prevalence and characterization of hippurate-negative Campylobacter jejuni in King County, Washington , 1987, Journal of clinical microbiology.

[14]  A. Simor,et al.  Evaluation of a blood-free, charcoal-based, selective medium for the isolation of Campylobacter organisms from feces , 1986, Journal of clinical microbiology.

[15]  G. Ederer,et al.  Rapid Hippurate Hydrolysis Method for Presumptive Identification of Group B Streptococci , 1975, Journal of clinical microbiology.

[16]  J. Butzler,et al.  Campylobacter: pathogenicity and significance in foods. , 1991, International journal of food microbiology.