Abstract
We describe a simple, rapid, and discriminatory methodology that allows the routine molecular characterization of Campylobacter jejuni and Campylobacter coli isolates. The proposed approach is built on one of the earliest and simplest molecular typing methods ever, consisting on the analysis of the fragments of different lengths generated by digestion of homologous DNA sequences with specific restriction endonucleases, a process known as restriction fragment length polymorphism (RFLP) analysis. The strategy underneath the workflow reported here is meant to explore the polymorphisms of Campylobacter spp. flaA gene (flaA-RFLP) that allows the local investigation of the genetic diversity and distribution of C. coli and C. jejuni isolates from different sources, namely, chickens’ caeca. Although not appropriate for global and long-term epidemiological studies as a single approach, flaA-RFLP analysis can be very useful in surveys limited in space and time and, for specific epidemiological settings, an alternative to more modern and resource-demanding techniques.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
References
FAO/WHO [Food and Agriculture Organization of the United Nations/World Health Organization] (2009) Risk assessment of Campylobacter spp. in broiler chickens: Technical Report. Microbiological Risk Assessment Series No 12. Geneva. pp 132
Miller WG, Mandrell RE (2005) Prevalence of Campylobacter in the food and water supply: incidence, outbreaks, isolation and detection. In: Ketley JM, Konkel M (eds) Campylobacter: molecular and cellular biology. Horizon Bioscience, Norfolk, UK, pp 101–108
Wilson DJ, Gabriel E, Leatherbarrow AJH et al (2008) Tracing the source of campylobacteriosis. PLoS Genet. doi:10.1371/journal.pgen.1000203
Wingstrand A, Neimann J, Engberg J et al (2006) Fresh chicken as main risk factor for campylobacteriosis, Denmark. Emerg Infect Dis 12:280–285
Humphrey T, O’Brien SJ, Madsen M (2007) Campylobacters as zoonotic pathogens: a food production perspective. Int J Food Microbiol 117:237–257
Dasti JI, Tareen AM, Raimond L et al (2010) Campylobacter jejuni: a brief overview on pathogenicity associated factors and disease-mediating mechanisms. Int J Med Microbiol 300:205–211
Silva J, Leite D, Fernandes M et al (2011) Campylobacter spp. as a foodborne pathogen: a review. Front Microbiol 2:1–12
Wassenaar T, Newell D (2000) Genotyping of Campylobacter spp. Appl Environ Microbiol 66:1–9
Klena JD, Konkel E (2005) Methods for epidemiological analysis of Campylobacter jejuni. In: Ketley JM, Konkel M (eds) Campylobacter: molecular and cellular biology. Horizon Bioscience, Norfolk, UK, pp 166–175
Fitzgerald C, Sails AD, Fields PI (2005) Campylobacter jejuni strain variation. In: Ketley JM, Konkel M (eds) Campylobacter: molecular and cellular biology. Horizon Bioscience, Norfolk, UK, pp 59–72
Jagannathan A, Penn C (2005) Motility. In: Ketley JM, Konkel M (eds) Campylobacter: molecular and cellular biology. Horizon Bioscience, Norfolk, UK, pp 331–344
Harrington CS, Thomson-Carter FM, Carter PE (1997) Evidence for recombination in the flagellin locus of Campylobacter jejuni; implications for the flagellin gene typing scheme. J Clin Microbiol 35:2836–2892
Carreira AC, Clemente L, Rocha T et al (2012) Comparative genotypic and antimicrobial susceptibility analysis of zoonotic Campylobacter species isolated from broilers on a nationwide survey, Portugal. J Food Prot 75:2100–2109
Ertas HB, Cetinkaya B, Muz A et al (2004) Genotyping of broiler-originated Campylobacter jejuni and Campylobacter coli isolates using fla typing and random amplified polymorphic DNA methods. Int J Food Microbiol 94:203–209
Nachamkin I, Bohachick K, Patton CM (1993) Flagellin gene typing of Campylobacter jejuni by restriction fragment length polymorphism analysis. J Clin Microbiol 31:1531–1536
Nachamkin I, Ung H, Patton CM (1996) Analysis of HL and O serotypes of Campylobacter strains by the flagellin gene typing system. J Clin Microbiol 34:277–281
Anonymous (2006) Microbiology of food and animal feeding stuffs—horizontal method for detection and enumeration of Campylobacter spp. Part 1. Detection method. ISO10272- 1:2006. International Organization for Standardization, Geneva
Hunter PR (1990) Reproducibility and indices of discriminatory power of microbial typing methods. J Clin Microbiol 28:1903–1905
Denis M, Soumet C, Rival K et al (1999) Development of a m-PCR assay for simultaneous identification of Campylobacter jejuni and Campylobacter coli. Lett Appl Microbiol 29:406–410
Wang G, Clark C, Taylor T et al (2002) Colony multiplex PCR assay for identification and differentiation of Campylobacter jejuni, C. coli, C. upsaliensis, and C. fetus subsp fetus. J Clin Microbiol 40:4744–4747
Author information
Authors and Affiliations
Corresponding author
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2015 Springer Science+Business Media New York
About this protocol
Cite this protocol
Carreira, A.C., Cunha, M.V. (2015). Characterization of Campylobacter jejuni and Campylobacter coli Genotypes in Poultry Flocks by Restriction Fragment Length Polymorphism (RFLP) Analysis. In: Cunha, M., Inácio, J. (eds) Veterinary Infection Biology: Molecular Diagnostics and High-Throughput Strategies. Methods in Molecular Biology, vol 1247. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-2004-4_22
Download citation
DOI: https://doi.org/10.1007/978-1-4939-2004-4_22
Published:
Publisher Name: Humana Press, New York, NY
Print ISBN: 978-1-4939-2003-7
Online ISBN: 978-1-4939-2004-4
eBook Packages: Springer Protocols