Methods ARTICLE

Front. Cell. Infect. Microbiol., 02 April 2012 | doi: 10.3389/fcimb.2012.00045

Multilocus sequence typing methods for the emerging Campylobacter species C. hyointestinalis, C. lanienae, C. sputorum, C. concisus, and C. curvus

William G. Miller1*, Mary H. Chapman1, Emma Yee1, Stephen L. W. On2, Desmond K. McNulty3, Albert J. Lastovica4, Anne M. Carroll3, Eleanor B. McNamara3, Geraldine Duffy5 and Robert E. Mandrell1
  • 1 USDA, ARS, WRRC, Produce Safety and Microbiology, Albany, CA, USA
  • 2 Christchurch Science Centre, Institute of Environmental Science and Research Limited, Christchurch, New Zealand
  • 3 Public Health Laboratory, Health Service Executive, Cherry Orchard Hospital, Ballyfermot, Dublin, Ireland
  • 4 Department of Biotechnology, University of the Western Cape, Bellville, South Africa
  • 5 Food Safety Department, Teagasc Food Research Centre, Ashtown, Dublin, Ireland

Multilocus sequence typing (MLST) systems have been reported previously for multiple food- and food animal-associated Campylobacter species (e.g., C. jejuni, C. coli, C. lari, and C. fetus) to both differentiate strains and identify clonal lineages. These MLST methods focused primarily on campylobacters of human clinical (e.g., C. jejuni) or veterinary (e.g., C. fetus) relevance. However, other, emerging, Campylobacter species have been isolated increasingly from environmental, food animal, or human clinical samples. We describe herein four MLST methods for five emerging Campylobacter species: C. hyointestinalis, C. lanienae, C. sputorum, C. concisus, and C. curvus. The concisus/curvus method uses the loci aspA, atpA, glnA, gltA, glyA, ilvD, and pgm, whereas the other methods use the seven loci defined for C. jejuni (i.e., aspA, atpA, glnA, gltA, glyA, pgm, and tkt). Multiple food animal and human clinical C. hyointestinalis (n = 48), C. lanienae (n = 34), and C. sputorum (n = 24) isolates were typed, along with 86 human clinical C. concisus and C. curvus isolates. A large number of sequence types were identified using all four MLST methods. Additionally, these methods speciated unequivocally isolates that had been typed ambiguously using other molecular-based speciation methods, such as 16S rDNA sequencing. Finally, the design of degenerate primer pairs for some methods permitted the typing of related species; for example, the C. hyointestinalis primer pairs could be used to type C. fetus strains. Therefore, these novel Campylobacter MLST methods will prove useful in differentiating strains of multiple, emerging Campylobacter species.

Keywords: MLST, emerging, Campylobacter hyointestinalis, Campylobacter lanienae, Campylobacter concisus, Campylobacter curvus, Campylobacter sputorum

Citation: Miller WG, Chapman MH, Yee E, On SLW, McNulty DK, Lastovica AJ, Carroll AM, McNamara EB, Duffy G and Mandrell RE (2012) Multilocus sequence typing methods for the emerging Campylobacter species C. hyointestinalis, C. lanienae, C. sputorum, C. concisus, and C. curvus. Front. Cell. Inf. Microbio. 2:45. doi: 10.3389/fcimb.2012.00045

Received: 05 December 2011; Accepted: 16 March 2012;
Published online: 02 April 2012.

Edited by:

Alain Stintzi, Ottawa Institute of Systems Biology, Canada

Reviewed by:

Qijing Zhang, Iowa State University, USA
Eduardo Taboada, Public Health Agency of Canada, Canada

Copyright: © 2012 Miller, Chapman, Yee, On, McNulty, Lastovica, Carroll, McNamara, Duffy and Mandrell. This is an open-access article distributed under the terms of the Creative Commons Attribution Non Commercial License, which permits non-commercial use, distribution, and reproduction in other forums, provided the original authors and source are credited.

*Correspondence: William G. Miller, USDA, ARS, WRRC, Produce Safety and Microbiology Research Unit, 800 Buchanan Street, Albany, CA 94710, USA. e-mail: william.miller@ars.usda.gov

Present address: Desmond K. McNulty, ICON Central Laboratories, South County Business Park, Leopardstown, Dublin, Ireland.

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