This Article Full Text (PDF) Alert me when this article is cited Alert me if a correction is posted Services Similar articles in this journal Similar articles in PubMed Alert me to new issues of the journal Download to citation manager Reprints and Permissions Copyright Information Books from ASM Press MicrobeWorld Citing Articles Citing Articles via HighWire Citing Articles via Google Scholar Google Scholar Articles by Giesendorf, B A Articles by Quint, W G Search for Related Content PubMed PubMed Citation Articles by Giesendorf, B A Articles by Quint, W G

Development of species-specific DNA probes for Campylobacter jejuni, Campylobacter coli, and Campylobacter lari by polymerase chain reaction fingerprinting.

Department of Molecular Biology, Diagnostic Center SSDZ, Delft, The Netherlands.

ABSTRACT

The application of polymerase chain reaction (PCR) fingerprinting assays enables discrimination between species and strains of microorganisms. PCR primers aiming at arbitrary sequences in combination with primers directed against the repetitive extragenic palindrome (REP) or enterobacterial repetitive intergenic consensus (ERIC) motifs generate isolate-specific DNA banding patterns. Analysis of these PCR fingerprints obtained for 33 isolates of Campylobacter jejuni, 30 isolates of Campylobacter coli, and 8 isolates of Campylobacter lari revealed that besides generation of isolate-specific fragments, species-specific DNA fragments of identical size were synthesized. It appeared that these DNA fragments could be used as species-specific probes, since they are unique for the pattern which they are deriving from. The probes do not cross-react with amplified DNA originating from a large panel of nonrelated microorganisms. Moreover, these probes displayed species specificity, as they reacted with a single restriction fragment on Southern blots containing DNA from C. jejuni, C. coli, and C. lari and other Campylobacter species. This combination of PCR fingerprinting and probe hybridization results in a highly specific identification assay and provides an example of specific test development without the prior need for DNA sequence information. The principle of the procedure holds great promise for the rapid isolation of DNA probes which, in combination with a general PCR assay, may lead to efficient typing and detection procedures for a multitude of medically important nonviral microorganisms.

This article has been cited by other articles:

• Moreno, Y., Botella, S., Alonso, J. L., Ferrus, M. A., Hernandez, M., Hernandez, J. (2003). Specific Detection of Arcobacter and Campylobacter Strains in Water and Sewage by PCR and Fluorescent In Situ Hybridization. Appl. Environ. Microbiol. 69: 1181-1186 [Abstract] [Full Text]  
• (2002). Molecular typing of micro-organisms: at the centre of diagnostics, genomics and pathogenesis of infectious diseases?. J Med Microbiol 51: 7-10 [Full Text]  
• Johnson, J. R., Clabots, C. (2000). Improved Repetitive-Element PCR Fingerprinting of Salmonella enterica with the Use of Extremely Elevated Annealing Temperatures. CVI 7: 258-264 [Abstract] [Full Text]  
• van Doorn, L.-J., Verschuuren-van Haperen, A., Burnens, A., Huysmans, M., Vandamme, P., Giesendorf, B. A. J., Blaser, M. J., Quint, W. G. V. (1999). Rapid Identification of Thermotolerant Campylobacter jejuni, Campylobacter coli, Campylobacter lari, and Campylobacter upsaliensis from Various Geographic Locations by a GTPase-Based PCR-Reverse Hybridization Assay. J. Clin. Microbiol. 37: 1790-1796 [Abstract] [Full Text]  
• van Belkum, A., Scherer, S., van Alphen, L., Verbrugh, H. (1998). Short-Sequence DNA Repeats in Prokaryotic Genomes. Microbiol. Mol. Biol. Rev. 62: 275-293 [Abstract] [Full Text]  
• Koeuth, T, Versalovic, J, Lupski, J R (1995). Differential subsequence conservation of interspersed repetitive Streptococcus pneumoniae BOX elements in diverse bacteria.. Genome Res. 5: 408-418 [Abstract]