Campylobacter fetus subspecies specific PCR assays inferred from comparative genomic analysis for accurate subspecies identification
- PMID: 39343039
- DOI: 10.1016/j.mimet.2024.107049
Campylobacter fetus subspecies specific PCR assays inferred from comparative genomic analysis for accurate subspecies identification
Abstract
Bovine Genital Campylobacteriosis (BGC) is caused by Campylobacter fetus subsp. venerealis and is a notifiable disease to the WOAH (World Organisation for Animal Health). For an effective BGC control program, the reliable differentiation of Campylobacter fetus subsp. venerealis (Cfv) from the closely related Campylobacter fetus subsp. fetus (Cff) is required. However, the available molecular C. fetus subspecies identification assays lack sensitivity and specificity to differentiate C. fetus isolates based on their phenotypic or genotypic differences. Furthermore, the current biochemical subspecies identification is not fully congruent with the genomic differentiation of C. fetus strains. In this study, the genome sequences of 41C. fetus strains with well identified subspecies, were analyzed with the large-scale BLAST score ratio (LS-BSR) pipeline to identify Cff and Cfv specific sequences. With this analysis, the asd gene encoding an aspartate-semialdehyde dehydrogenase was identified, which contained a 6-bp Cff-specific sequence, and this 6-bp sequence was absent in the asd gene of Cfv strains. This sequence was used for the development of PCR assays to differentiate Cff and Cfv strains. The C. fetus subspecies identification of the developed asd PCR assays was in full congruence with the genomic classification of strains and are recommended for molecular identification of C. fetus subspecies in BGC control programs. The asd PCR can be assessed on sequenced genomes using a web interface containing the Cfvcatch tool, which includes placement of the tested genome in a phylogenetic tree with reference C. fetus genomes to distinguish the two subspecies and to detect antimicrobial resistance genes.
Keywords: Bovine Genital Campylobacteriosis; Campylobacter fetus; Campylobacter fetus subspecies venerealis; Subspecies identification.
Copyright © 2024 The Authors. Published by Elsevier B.V. All rights reserved.
Conflict of interest statement
Declaration of competing interest The author(s) declare that there are no conflicts of interest.
Similar articles
-
Development of loop-mediated isothermal amplification and PCR assays for rapid and simple detection of Campylobacter fetus subsp. venerealis.Microbiol Immunol. 2010 Jul;54(7):398-404. doi: 10.1111/j.1348-0421.2010.00233.x. Microbiol Immunol. 2010. PMID: 20618686
-
Genomic analysis of Campylobacter fetus subspecies: identification of candidate virulence determinants and diagnostic assay targets.BMC Microbiol. 2009 May 8;9:86. doi: 10.1186/1471-2180-9-86. BMC Microbiol. 2009. PMID: 19422718 Free PMC article.
-
Evaluation of molecular assays for identification Campylobacter fetus species and subspecies and development of a C. fetus specific real-time PCR assay.J Microbiol Methods. 2013 Oct;95(1):93-7. doi: 10.1016/j.mimet.2013.06.005. Epub 2013 Jun 26. J Microbiol Methods. 2013. PMID: 23810970
-
Assessment of Campylobacter fetus subsp. venerealis molecular diagnosis using clinical samples of bulls.BMC Vet Res. 2020 Oct 29;16(1):410. doi: 10.1186/s12917-020-02634-7. BMC Vet Res. 2020. PMID: 33121492 Free PMC article.
-
So close and yet so far - Molecular Microbiology of Campylobacter fetus subspecies.Eur J Microbiol Immunol (Bp). 2012 Mar;2(1):66-75. doi: 10.1556/EuJMI.2.2012.1.10. Epub 2012 Mar 17. Eur J Microbiol Immunol (Bp). 2012. PMID: 24611123 Free PMC article. Review.
MeSH terms
Substances
LinkOut - more resources
Full Text Sources
Medical
Research Materials