Exploiting bacterial whole-genome sequencing data for the evaluation of diagnostic assays: Campylobacter speciation as a case study


Van Rensburg MJJ, Swift C, Cody AJ, Jenkins C, Maiden MCJ

Unit Authors:

Martin Maiden, Melissa Jansen van Rensburg


The application of whole-genome sequencing (WGS) to problems in clinical microbiology has had a major impact on the field. Clinical laboratories are now using WGS for pathogen identification, antimicrobial susceptibility testing, and epidemiological typing. WGS data also represents a valuable resource for the development and evaluation of molecular diagnostic assays, which continue to play an important role in clinical microbiology. To demonstrate this application of WGS, the current study used publicly available genomic data to evaluate a duplex real-time PCR (RT-PCR) assay that targets mapA and ceuE for the detection of Campylobacter jejuni and Campylobacter coli, leading global causes of bacterial gastroenteritis. In silico analyses of mapA and ceuE primer and probe sequences from 1,713 genetically diverse C. jejuni and C. Coli genomes, supported by RT-PCR testing, indicated that the assay was robust, with 1,707 (99.7%) isolates correctly identified. The high specificity of the mapA/ceuE assay was the result of interspecies diversity and intraspecies conservation of the target genes in C. Jejuni and C. coli. Rare instances of a lack of specificity among C. coli isolates were due to introgression in mapA or sequence diversity in ceuE. The results of this study illustrate how WGS can be exploited to evaluate molecular diagnostic assays using publicly available data, online databases, and open source software.


Journal of Clinical Microbiology





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