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Journal of Clinical Microbiology, September 2003, p. 4071-4080, Vol. 41, No. 9
0095-1137/03/$08.00+0     DOI: 10.1128/JCM.41.9.4071-4080.2003

Microarray-Based Identification of Thermophilic Campylobacter jejuni, C. coli, C. lari, and C. upsaliensis

Dmitriy Volokhov,1 Vladimir Chizhikov,2 Konstantin Chumakov,2 and Avraham Rasooly1*

FDA Center for Food Safety and Applied Nutrition, College Park,1 FDA Center for Biologics Evaluation and Research, Rockville,Maryland2

Received 9 December 2002/ Returned for modification 3 April 2003/ Accepted 25 June 2003

DNA microarrays are an excellent potential tool for clinical microbiology, since this technology allows relatively rapid identification and characterization of microbial and viral pathogens. In the present study, an oligonucleotide microarray was developed and used for the analysis of thermophilic Campylobacter spp., the primary food-borne pathogen in the United States. We analyzed four Campylobacter species: Campylobacter jejuni, C. coli, C. lari, and C. upsaliensis. Our assay relies on the PCR amplification of specific regions in five target genes (fur, glyA, cdtABC, ceuB-C, and fliY) as a first step, followed by microarray-based analysis of amplified DNAs. Alleles of two genes, fur and glyA, which are found in all tested thermophilic Campylobacter spp., were used for identification and discrimination among four bacterial species, the ceuB-C gene was used for discrimination between C. jejuni and C. coli, and the fliY and cdt genes were used as additional genetic markers specific either for C. upsaliensis and C. lari or for C. jejuni. The array was developed and validated by using 51 previously characterized Campylobacter isolates. All isolates were unambiguously identified on the basis of hybridization patterns with 72 individual species-specific oligoprobes. Microarray identification of C. jejuni and C. coli was confirmed by PCR amplification of other genes used for identification (hipO and ask). Our results demonstrate that oligonucleotide microarrays are suitable for rapid and accurate simultaneous differentiation among C. jejuni, C. coli, C. lari, and C. upsaliensis.

* Corresponding author. Mailing address: Food and Drug Administration, 5100 Paint Branch Pkwy., College Park, MD 20740-3835. Phone: (301) 436-2016. Fax: (301) 436-2644. E-mail: axr{at}cfsan.fda.gov.

Journal of Clinical Microbiology, September 2003, p. 4071-4080, Vol. 41, No. 9
0095-1137/03/$08.00+0     DOI: 10.1128/JCM.41.9.4071-4080.2003




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