Abstract
Helicobacter pylori is an important pathogen whose primary niche is the human stomach. H. pylori is etiologically associated with gastric inflammation (gastritis), peptic ulcer disease, and gastric cancer. Both noninvasive (e.g., urea breath and stool antigen tests) and invasive (gastric biopsy for histology, culture, or PCR) tests are used for diagnosis. PCR detection of H. pylori has been reported using a variety of clinical samples including gastric biopsy, gastric juice, saliva, dental plaque, and stools as well as environmental samples. Whenever possibly, noninvasive tests are preferred over invasive tests. H. pylori are excreted in the stool. Culture from stool is variable whereas stool antigen testing is widely used. Stool consists of a complicated mixture of commensal bacteria and chemicals and often includes inhibitors of PCR. Nevertheless, simple extraction methods are available to efficiently extract DNA from human stools and nested-PCR targeting the 23S rRNA gene have proven to be highly sensitive for the detection of H. pylori. Detection of clarithromycin susceptibility/resistance is important clinically and the mutation of the 23S rRNA gene responsible for resistance can also be detected using stool. This described method can be modified for other clinical samples such as gastric juice or biopsy material.
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Acknowledgments
Dr. Rimbara and Dr. Sasatsu are supported by a grant from the High-Tech Research Centre Project for Private Universities provided by the Ministry of Education, Culture, Sports, Science and Technology and by the Matching Fund Subsidy for Private Schools of Japan. This work was supported in part by the Office of Research and Development Medical Research Service Department of Veterans Affairs. Dr. Graham is supported in part by Public Health Service grant DK56338 which funds the Texas Medical Center Digestive Diseases Center and R01 CA116845. We especially thank Drs. Norihisa Noguchi, Prof. Takashi Kawai, and Prof. Shinichi Takahashi for their expertise, advice, and encouragement.
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Rimbara, E., Sasatsu, M., Graham, D.Y. (2013). PCR Detection of Helicobacter pylori in Clinical Samples. In: Wilks, M. (eds) PCR Detection of Microbial Pathogens. Methods in Molecular Biology, vol 943. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-60327-353-4_19
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DOI: https://doi.org/10.1007/978-1-60327-353-4_19
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