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Use of an improved atpA amplification and sequencing method to identify members of the C ampylobacteraceae and H elicobacteraceae
Author(s) -
Miller W.G.,
Yee E.,
Jolley K.A.,
Chapman M.H.
Publication year - 2014
Publication title -
letters in applied microbiology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.698
H-Index - 110
eISSN - 1472-765X
pISSN - 0266-8254
DOI - 10.1111/lam.12228
Subject(s) - computational biology , biology , dna sequencing , genetics , gene
Emerging C ampylobacter and A rcobacter spp. have been increasingly isolated from human clinical samples, food, veterinary samples and the environment. Unambiguous species identification of such organisms is of obvious importance in epidemiological studies, but is also necessary to accurately assess their host range and determine their prevalence in the food chain and in the environment. Species identification methods for the C ampylobacteraceae have been described; however, some with high resolving power are limited to a small number of taxa, while other broader‐range methods cannot distinguish between closely related species. We present in this study a novel species identification method, based on amplification and sequencing of a portion of the atpA gene. This method, which uses a single primer pair, was able to amplify and accurately identify all current taxa within C ampylobacter and A rcobacter as well as several members of the H elicobacteraceae, although unambiguous identification of the C amp. fetus subspecies could not be achieved. In addition, five putative novel C ampylobacter taxa were recognized, making this new species identification method valuable in the characterization of novel epsilonproteobacteria. Thus, a single‐locus method that can accurately identify multiple epsilonproteobacterial species will prove important in the characterization of emerging organisms and those associated with illness. Significance and Impact of the Study The atpA ‐based species identification method described here uses a single primer pair to amplify DNA from all current validly‐described C ampylobacter and A rcobacter taxa, as well as multiple members of the H elicobacteraceae. This method unambiguously identified all taxa tested, although it could not discriminate the subspecies of C amp. fetus . Furthermore, five putative novel C ampylobacter taxa were observed following testing of environmental campylobacters with this method. The scope and resolution of this method make it an important addition to studies of epsilonproteobacterial epidemiology and evolution.