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United States Department of Agriculture

Agricultural Research Service

Research Project: Molecular Biology of Human Pathogens Associated with Food

Location: Produce Safety and Microbiology Research

Title: Use of an improved atpA amplification and sequencing method to identify members of the Campylobacteraceae and Helicobacteraceae

Authors
item Miller, William
item Yee, Emma
item Jolley, Keith -
item Chapman, Mary

Submitted to: Letters in Applied Microbiology
Publication Type: Peer Reviewed Journal
Publication Acceptance Date: February 5, 2014
Publication Date: March 8, 2014
Citation: Miller, W.G., Yee, E., Jolley, K.A., Chapman, M.H. 2014. Use of an improved atpA amplification and sequencing method to identify members of the Campylobacteraceae and Helicobacteraceae. Letters in Applied Microbiology. 58(6):582-590. DOI: 10.1111/lam.12228.

Interpretive Summary: Campylobacter have been isolated from a wide variety of environments, warm-blooded animals (birds, mammals) generally, but also occasionally cold-blooded reptiles. Campylobacters cause disease in both livestock and humans; human disease is primarily gastroenteritis, however other clinical outcomes, such as septicemia, can occur. Other related organisms such as arcobacters and helicobacters are also associated with human disease. Although the primary pathogens in this group are C. jejuni and H. pylori (the organism associated with ulcers and gastric cancer), other “emerging” campylobacters and arcobacters are increasingly associated with disease. There is a need for a rapid, unambiguous typing method to identify these emerging organisms. This study describes a new typing method that accurately identifies all campylobacters and arcobacters and multiple helicobacters, including H. pylori. This method can be used to type these potential pathogens in order to better understand their prevalence in the environment and in the food chain, and their association with human disease.

Technical Abstract: Emerging Campylobacter spp. and arcobacters have been increasingly isolated from human clinical samples, food 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 Campylobacteraceae have been described; however, some with high resolving power are limited to a small number of taxa, while other, broader-range methods cannot, e.g., 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 Campylobacter and Arcobacter as well as several members of the Helicobacteraceae, although unambiguous identification of the C. fetus subspecies could not be achieved. In addition, five putative novel Campylobacter taxa were identified, making this new typing method valuable in the characterization of novel epsilonproteobacteria. Thus, a single locus typing method that can accurately identify multiple epsilonproteobacterial species will prove important in the characterization of both emerging organisms and those currently associated with human illness.

Last Modified: 10/1/2014
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