AUTHOR=Miller William G., Chapman Mary H., Yee Emma , On Stephen L., McNulty Desmond K., Lastovica Albert J., Carroll Anne M., McNamara Eleanor B., Duffy Geraldine , Mandrell Robert E.

TITLE=Multilocus Sequence Typing Methods for the Emerging Campylobacter Species C. hyointestinalis, C. lanienae, C. sputorum, C. concisus, and C. curvus

JOURNAL=Frontiers in Cellular and Infection Microbiology

VOLUME=2

YEAR=2012

URL=https://www.frontiersin.org/journals/cellular-and-infection-microbiology/articles/10.3389/fcimb.2012.00045

DOI=10.3389/fcimb.2012.00045

ISSN=2235-2988

ABSTRACT=<p>Multilocus sequence typing (MLST) systems have been reported previously for multiple food- and food animal-associated <italic>Campylobacter</italic> species (e.g., <italic>C. jejuni</italic>, <italic>C. coli</italic>, <italic>C. lari</italic>, and <italic>C. fetus</italic>) to both differentiate strains and identify clonal lineages. These MLST methods focused primarily on campylobacters of human clinical (e.g., <italic>C. jejuni</italic>) or veterinary (e.g., <italic>C. fetus</italic>) relevance. However, other, emerging, <italic>Campylobacter</italic> species have been isolated increasingly from environmental, food animal, or human clinical samples. We describe herein four MLST methods for five emerging <italic>Campylobacter</italic> species: <italic>C. hyointestinalis</italic>, <italic>C. lanienae</italic>, <italic>C. sputorum</italic>, <italic>C. concisus</italic>, and <italic>C. curvus</italic>. The <italic>concisus</italic>/<italic>curvus</italic> method uses the loci <italic>aspA</italic>, <italic>atpA</italic>, <italic>glnA</italic>, <italic>gltA</italic>, <italic>glyA</italic>, <italic>ilvD</italic>, and <italic>pgm</italic>, whereas the other methods use the seven loci defined for <italic>C. jejuni</italic> (i.e., <italic>aspA</italic>, <italic>atpA</italic>, <italic>glnA</italic>, <italic>gltA</italic>, <italic>glyA</italic>, <italic>pgm</italic>, and <italic>tkt</italic>). Multiple food animal and human clinical <italic>C. hyointestinalis</italic> (<italic>n</italic> = 48), <italic>C. lanienae</italic> (<italic>n</italic> = 34), and <italic>C. sputorum</italic> (<italic>n</italic> = 24) isolates were typed, along with 86 human clinical <italic>C. concisus</italic> and <italic>C. curvus</italic> 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 <italic>C. hyointestinalis</italic> primer pairs could be used to type <italic>C. fetus</italic> strains. Therefore, these novel <italic>Campylobacter</italic> MLST methods will prove useful in differentiating strains of multiple, emerging <italic>Campylobacter</italic> species.</p>