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Multiplex Accelerated PCR System for One-Step Helicobacter pylori cagA + Genotypes Detection: A Guide for Clinical Testing

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Abstract

Helicobacter pylori cagA + genotype is a leading risk factor for gastric cancer development making accurate identification and timely eradication of H. pylori critical to deadly gastric cancer prevention. Traditional clinical diagnostic methods, including conventional in vitro culture, histological examination, and (13/14)C-urea breath test methods, could only identify the presence of H. pylori, but these means are not capable of identification of cagA + strains. Herein, we firstly built a multiplex detection system based on novel accelerated PCR that could realize one-step detection of as low as 20 copies of H. pylori 16S rDNA and cagA genes within 30 min. In addition, this novel system performed strong anti-jamming capacity, and exhibited that it could specifically differentiate H. pylori cagA- and cagA + genotypes co-existence with other 4 kinds of gastrointestinal pathogens. Furthermore, this one-step system showed remarkable performance on rapid H. pylori infection diagnosis and cagA + genotypes identification in clinical gastric mucosa samples. Specifically, it outperformed histological examination in terms of accuracy and was superior to conventional PCR and DNA sequencing in terms of efficiency. This rapid, sensitive, and reliable H. pylori detection and identification system would break the limitation of traditional methods and realize H. pylori infection diagnosis and cagA + genotypes identification.

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Acknowledgements

This work was financially supported by the National Key Research and Development Programs of China (2018YFE0113300) and the Key Project of Shandong Province Natural Science Foundation (ZR2020KH030), as well as the clinical samples donated by the Affiliated Hospital of Qingdao University.

Funding

This work was financially supported by the National Key Research and Development Programs of China (2018YFE0113300) and the Key Project of Shandong Province Natural Science Foundation (ZR2020KH030).

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Author notes

  1. Yanling Wang, Yang Li these two authors contributed equally to this work.

Authors and Affiliations

  1. Qingdao Nucleic Acid Rapid Testing International Science and Technology Cooperation Base, College of Life Sciences, Department of Pathogenic Biology, School of Basic Medicine, and Department of Clinical Laboratory, The Affiliated Hospital of Qingdao University, Qingdao University, Qingdao, 266071, Shandong, China

    Yanling Wang, Yang Li, Zhixian Luan, Yan Zhao, Peng Zhang & Chao Shi

  2. Key Laboratory of Optic-Electric Sensing and Analytical Chemistry for Life Science, MOE,, Shandong Provincial Key Laboratory of Biochemical Engineering, Qingdao Nucleic Acid Rapid Detection Engineering Research Center, College of Marine Science and Biological Engineering, Qingdao University of Science and Technology, Qingdao, 266042, People’s Republic of China

    Cuiping Ma

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  1. Yanling Wang
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Contributions

YW, ZL, and PZ performed the experiments; YZ collected the gastric mucosa samples; YL and YW analyzed the data; YL, CM, and CS designed the study; YL and YW wrote the manuscript; and all authors contributed to the writing of the paper, had primary responsibility for the final content, and read and approved the final manuscript.

Corresponding author

Correspondence to Chao Shi.

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There are no conflicts of interest to declare.

Ethical Approval

All sample collection work and tests were conducted with the patients’ consent and approved by the authorized Human Health and Ethics Committee of the Affiliated Hospital of Qingdao University (Approval No. QDU-HEC-2021137), and all operations were conducted following relevant guidelines and regulations.

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Wang, Y., Li, Y., Luan, Z. et al. Multiplex Accelerated PCR System for One-Step Helicobacter pylori cagA + Genotypes Detection: A Guide for Clinical Testing. Curr Microbiol 79, 235 (2022). https://doi.org/10.1007/s00284-022-02931-4

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