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Real-time single-nucleotide polymorphism profiling using Taqman technology for rapid recognition of Campylobacter jejuni clonal complexes

E L Best¹, A J Fox², J A Frost¹ and F J Bolton²

¹Campylobacter and Helicobacter Reference Unit, Laboratory of Enteric Pathogens, Centre For Infections, Health Protection Agency, 61 Colindale Avenue, London NW9 5HT, UK ²Health Protection Agency North West Laboratory, Manchester Medical Microbiology Partnership, Manchester Royal Infirmary, Manchester, UK

Correspondence E. L. Best emma.best{at}hpa.org.uk

Received December 2, 2004
Accepted June 22, 2005

The rapid identification of Campylobacter jejuni isolates to strain level would significantly inform the public health investigation of C. jejuni infection. Conceptual advances provided by multilocus sequence typing (MLST) have established the clonal complex as an important epidemiological group at the strain level, enabling accurate and phylogenetically valid strain identification for C. jejuni. The development of real-time PCR assays for allelic discrimination of strain-associated single-nucleotide polymorphisms (SNPs) based upon MLST locus alleles offers one possible approach for rapid strain detection. SNPs defining key alleles diagnostic for the most prevalent clonal complexes were identified following a detailed analysis of the available MLST data. Real-time Taqman allelic discrimination assays designed to detect the SNPs specific for six major clonal complexes, ST-21, ST-45, ST-48, ST-61, ST-206 and ST-257, were developed, allowing the rapid detection of C. jejuni isolates and preliminary strain identification. This will provide an important complementary technique to sequence typing for rapid detection and strain characterization to inform in real-time the public health management and investigation of C. jejuni infections.

Abbreviations: C_T, threshold cycle; MLST, multilocus sequence typing; SNP, single-nucleotide polymorphism.

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