Real-time detection of Campylobacter jejuni and Campylobacter coli from environmental samples and single nucleotide polymorphism profiling of map A positive strains to determine their clonal complexes
Abstract number: p1680
Lai S., Best E., Surman-Lee S.B., Owen R.J., Lee J.V.
A study to demonstrate the rapid detection and speciation of Campylobacter jejuni and of Campylobacter coli isolates directly from enrichment broth using a Taqman® assay. Single nucleotide polymorphism analysis of mapA positive strains was used for rapid identification of C. jejuni clonal complexes.
Thermotolerant Campylobacter species were initially confirmed by culture according to the modified draft ISO 17995 method, where water samples were filtered through 0.2 mm pore size nylon membrane. The filters were transferred to selective enrichment in Preston broth to improve their recovery and therefore detection of any campylobacter cells present. DNA was extracted directly from the enrichment broth culture for real-time detection of C. jejuni and C. coli using the Taqman®. Samples, which were map A positive were, further characterise by single nucleotide polymorphism profiling for rapid recognition of C. jejuni clonal complexes.
Environmental samples, which were confirmed by culture were also map A positive by Taqman®. SNP profiling of mapA positive isolates identified clonal complexes, which are predominantly contained in isolates of human disease and chicken.
This study has demonstrated the feasibility of rapid detection and identification of C. jejuni and C. coli following short enrichment incubation using a Taqman® assay. A rapid turnaround time of between 34 h per batch of 96 samples was achieved. SNP profiling offers important epidemiological grouping at strain level, enabling accurate and phylogenetically valid strain identification for C. jejuni, which may have important host associations for tracing sources of infection and consequently improve public health responses.
|Session name:||XXIst ISTH Congress|
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