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. 1992 Dec;58(12):3804–3808. doi: 10.1128/aem.58.12.3804-3808.1992

Rapid and sensitive detection of Campylobacter spp. in chicken products by using the polymerase chain reaction.

B A Giesendorf 1, W G Quint 1, M H Henkens 1, H Stegeman 1, F A Huf 1, H G Niesters 1
PMCID: PMC183185  PMID: 1282312

Abstract

The polymerase chain reaction (PCR) after a short enrichment culture was used to detect Campylobacter spp. in chicken products. After the 16S rRNA gene sequence of Campylobacter jejuni was determined and compared with known sequences from other enterobacteria, a primer and probe combination was selected from the region before V3 and the variable regions V3 and V5. With this primer set and probe, 426-bp fragments from C. jejuni, Campylobacter coli, and Campylobacter lari could be amplified. The detection limit of the PCR was 12.5 CFU. Chicken samples inoculated with 25 CFU of Campylobacter spp. per g were PCR positive after an 18-h enrichment, which resulted in 500 CFU/ml of culture broth. This PCR-culture assay was compared with the conventional method on naturally infected chicken products. Both methods detected the same number of positive and negative samples; however, the results of the PCR-culture assay were available within 48 h.

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Selected References

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  1. Blaser M. J., Reller L. B. Campylobacter enteritis. N Engl J Med. 1981 Dec 10;305(24):1444–1452. doi: 10.1056/NEJM198112103052404. [DOI] [PubMed] [Google Scholar]
  2. Boom R., Sol C. J., Salimans M. M., Jansen C. L., Wertheim-van Dillen P. M., van der Noordaa J. Rapid and simple method for purification of nucleic acids. J Clin Microbiol. 1990 Mar;28(3):495–503. doi: 10.1128/jcm.28.3.495-503.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Brosius J., Palmer M. L., Kennedy P. J., Noller H. F. Complete nucleotide sequence of a 16S ribosomal RNA gene from Escherichia coli. Proc Natl Acad Sci U S A. 1978 Oct;75(10):4801–4805. doi: 10.1073/pnas.75.10.4801. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Hill W. E., Keasler S. P., Trucksess M. W., Feng P., Kaysner C. A., Lampel K. A. Polymerase chain reaction identification of Vibrio vulnificus in artificially contaminated oysters. Appl Environ Microbiol. 1991 Mar;57(3):707–711. doi: 10.1128/aem.57.3.707-711.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Lampel K. A., Jagow J. A., Trucksess M., Hill W. E. Polymerase chain reaction for detection of invasive Shigella flexneri in food. Appl Environ Microbiol. 1990 Jun;56(6):1536–1540. doi: 10.1128/aem.56.6.1536-1540.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Neefs J. M., Van de Peer Y., Hendriks L., De Wachter R. Compilation of small ribosomal subunit RNA sequences. Nucleic Acids Res. 1990 Apr 25;18 (Suppl):2237–2317. doi: 10.1093/nar/18.suppl.2237. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Olive D. M., Johny M., Sethi S. K. Use of an alkaline phosphatase-labeled synthetic oligonucleotide probe for detection of Campylobacter jejuni and Campylobacter coli. J Clin Microbiol. 1990 Jul;28(7):1565–1569. doi: 10.1128/jcm.28.7.1565-1569.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Park R. W., Griffiths P. L., Moreno G. S. Sources and survival of campylobacters: relevance to enteritis and the food industry. Soc Appl Bacteriol Symp Ser. 1991;20:97S–106S. [PubMed] [Google Scholar]
  9. 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]
  10. Rollins D. M., Colwell R. R. Viable but nonculturable stage of Campylobacter jejuni and its role in survival in the natural aquatic environment. Appl Environ Microbiol. 1986 Sep;52(3):531–538. doi: 10.1128/aem.52.3.531-538.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Romaniuk P. J., Trust T. J. Rapid identification of Campylobacter species using oligonucleotide probes to 16S ribosomal RNA. Mol Cell Probes. 1989 Jun;3(2):133–142. doi: 10.1016/0890-8508(89)90024-8. [DOI] [PubMed] [Google Scholar]
  12. Romaniuk P. J., Zoltowska B., Trust T. J., Lane D. J., Olsen G. J., Pace N. R., Stahl D. A. Campylobacter pylori, the spiral bacterium associated with human gastritis, is not a true Campylobacter sp. J Bacteriol. 1987 May;169(5):2137–2141. doi: 10.1128/jb.169.5.2137-2141.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Saiki R. K., Gelfand D. H., Stoffel S., Scharf S. J., Higuchi R., Horn G. T., Mullis K. B., Erlich H. A. Primer-directed enzymatic amplification of DNA with a thermostable DNA polymerase. Science. 1988 Jan 29;239(4839):487–491. doi: 10.1126/science.2448875. [DOI] [PubMed] [Google Scholar]
  14. Skirrow M. B. Campylobacter. Lancet. 1990 Oct 13;336(8720):921–923. doi: 10.1016/0140-6736(90)92282-m. [DOI] [PubMed] [Google Scholar]
  15. Tenover F. C., Carlson L., Barbagallo S., Nachamkin I. DNA probe culture confirmation assay for identification of thermophilic Campylobacter species. J Clin Microbiol. 1990 Jun;28(6):1284–1287. doi: 10.1128/jcm.28.6.1284-1287.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Thorne G. M., Macone A., Goldmann D. A. Enzymatically labelled nucleic acid (NA) probe assays for detection of Campylobacter spp. in human faecal specimens and in culture. Mol Cell Probes. 1990 Apr;4(2):133–142. doi: 10.1016/0890-8508(90)90014-q. [DOI] [PubMed] [Google Scholar]
  17. Vandamme P., Falsen E., Rossau R., Hoste B., Segers P., Tytgat R., De Ley J. Revision of Campylobacter, Helicobacter, and Wolinella taxonomy: emendation of generic descriptions and proposal of Arcobacter gen. nov. Int J Syst Bacteriol. 1991 Jan;41(1):88–103. doi: 10.1099/00207713-41-1-88. [DOI] [PubMed] [Google Scholar]
  18. Walker R. I., Caldwell M. B., Lee E. C., Guerry P., Trust T. J., Ruiz-Palacios G. M. Pathophysiology of Campylobacter enteritis. Microbiol Rev. 1986 Mar;50(1):81–94. doi: 10.1128/mr.50.1.81-94.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Wernars K., Delfgou E., Soentoro P. S., Notermans S. Successful approach for detection of low numbers of enterotoxigenic Escherichia coli in minced meat by using the polymerase chain reaction. Appl Environ Microbiol. 1991 Jul;57(7):1914–1919. doi: 10.1128/aem.57.7.1914-1919.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Wilson I. G., Cooper J. E., Gilmour A. Detection of enterotoxigenic Staphylococcus aureus in dried skimmed milk: use of the polymerase chain reaction for amplification and detection of staphylococcal enterotoxin genes entB and entC1 and the thermonuclease gene nuc. Appl Environ Microbiol. 1991 Jun;57(6):1793–1798. doi: 10.1128/aem.57.6.1793-1798.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Wilson K. H., Blitchington R. B., Greene R. C. Amplification of bacterial 16S ribosomal DNA with polymerase chain reaction. J Clin Microbiol. 1990 Sep;28(9):1942–1946. doi: 10.1128/jcm.28.9.1942-1946.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]

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