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Journal of Clinical Microbiology, June 1999, p. 1790-1796, Vol. 37, No. 6
0095-1137/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.

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

Leen-Jan van Doorn,1,* Anita Verschuuren-van Haperen,1 André Burnens,2 Michael Huysmans,3 Peter Vandamme,4,5 Belinda A. J. Giesendorf,6 Martin J. Blaser,7 and Wim G. V. Quint1

Institute for Veterinary Bacteriology, University of Bern, Bern, Switzerland2; Department of Microbiology, Monash Medical Center, Clayton, Victoria, Australia3; Laboratory of Microbiology, University of Ghent, Ghent,4 and Department of Medical Microbiology, University Hospital UIA, Antwerp,5 Belgium; Delft Diagnostic Laboratory, Delft,1 and Department of Clinical Chemistry (CKCL), University Hospital Nijmegen, Nijmegen,6 The Netherlands; and Division of Infectious Diseases and VA Medical Center, Vanderbilt University, Nashville, Tennessee7

Received 3 November 1998/Returned for modification 2 February 1999/Accepted 4 March 1999

Recently, a gene from Campylobacter jejuni encoding a putative GTPase was identified. Based on two semiconserved GTP-binding sites encoded within this gene, PCR primers were selected that allow amplification of a 153-bp fragment from C. jejuni, C. coli, C. lari, and C. upsaliensis. Sequence analysis of these PCR products revealed consistent interspecies variation, which allowed the definition of species-specific probes for each of the four thermotolerant Campylobacter species. Multiple probes were used to develop a line probe assay (LiPA) that permits analysis of PCR products by a single reverse hybridization step. A total of 320 reference strains and clinical isolates from various geographic origins were tested by the GTP-based PCR-LiPA. The PCR-LiPA is highly specific in comparison with conventional identification methods, including biochemical and whole-cell protein analyses. In conclusion, a simple method has been developed for rapid and highly specific identification of thermotolerant Campylobacter species.

* Corresponding author. Mailing address: R. de Graafweg 7, 2625 AD, Delft, The Netherlands. Phone: 31-15 2604577. Fax: 31-15 2604550. E-mail: L.J.van.Doorn{at}ddl.nl.

Journal of Clinical Microbiology, June 1999, p. 1790-1796, Vol. 37, No. 6
0095-1137/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.


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