Basic Study
Copyright ©The Author(s) 2021. Published by Baishideng Publishing Group Inc. All rights reserved.
World J Gastroenterol. Jun 28, 2021; 27(24): 3595-3608
Published online Jun 28, 2021. doi: 10.3748/wjg.v27.i24.3595
Early genetic diagnosis of clarithromycin resistance in Helicobacter pylori
Xiao-Hua Li, Yong-Yi Huang, Lin-Ming Lu, Li-Juan Zhao, Xian-Ke Luo, Ru-Jia Li, Yuan-Yuan Dai, Chun Qin, Yan-Qiang Huang, Hao Chen
Xiao-Hua Li, Yong-Yi Huang, Li-Juan Zhao, Ru-Jia Li, Yuan-Yuan Dai, Chun Qin, Yan-Qiang Huang, Research Center for the Prevention and Treatment of Drug Resistant Microbial Infection, Youjiang Medical University for Nationalities, Baise 533000, Guangxi Zhuang Autonomous Region, China
Lin-Ming Lu, Hao Chen, Department of Pathology, Wannan Medical College, Wuhu 241002, Anhui Province, China
Xian-Ke Luo, Department of Gastroenterology, National Hospital of Guangxi Zhuang Autonomous Region, Nanning Guangxi Zhuang Autonomous Region, 530001, China
Author contributions: Li XH and Huang YY contributed equally to this work, and they consulted the literature, performed the experiments, acquired and analyzed the data, and wrote the first draft; Lu LM, Zhao LJ, Luo XK, Li RJ, Dai YY, and Qin C revised the manuscript; Huang YQ and Chen H served as corresponding authors, contributed equally to this work, contributed equally to this work, and they designed, checked, and revised the final manuscript; all authors approved the final version of the article.
Supported by National Natural Science Foundation of China, No. 81760739 and No. 31460023; and Special Fund Projects for Guiding Local Science and Technology Development by the Chinese Government, No. GUIKEZY20198004.
Institutional review board statement: The study was reviewed and approved by the Institutional Review Board at Youjiang Medical University for Nationalities.
Conflict-of-interest statement: Li XH, Huang YY, Zhao LJ, Li RJ, Dai YY, Qin C, and Huang YQ are employed by Youjiang Medical University for Nationalities; Lu LM and Chen H are employed by Wannan Medical College; Luo XK is employed by National Hospital of Guangxi Zhuang Autonomous Region; all other authors have nothing to disclose.
Data sharing statement: No additional data are available.
Open-Access: This article is an open-access article that was selected by an in-house editor and fully peer-reviewed by external reviewers. It is distributed in accordance with the Creative Commons Attribution NonCommercial (CC BY-NC 4.0) license, which permits others to distribute, remix, adapt, build upon this work non-commercially, and license their derivative works on different terms, provided the original work is properly cited and the use is non-commercial. See: http://creativecommons.org/Licenses/by-nc/4.0/
Corresponding author: Yan-Qiang Huang, MD, PhD, Professor, Research Center for the Prevention and Treatment of Drug Resistant Microbial Infection, Youjiang Medical University for Nationalities, No. 98 Countryside Road, Baise 533000, Guangxi Zhuang Autonomous Region, China, hyq77615@163.com
Received: March 2, 2021
Peer-review started: March 2, 2021
First decision: April 5, 2021
Revised: April 13, 2021
Accepted: May 21, 2021
Article in press: May 21, 2021
Published online: June 28, 2021
Processing time: 115 Days and 1 Hours
Abstract
BACKGROUND

The drug resistance rate of clinical Helicobacter pylori (H. pylori) isolates has increased. However, the mechanism of drug resistance remains unclear. In this study, drug-resistant H. pylori strains were isolated from different areas and different populations of Chinese for genomic analysis.

AIM

To investigate drug-resistant genes in H. pylori and find the genes for the early diagnosis of clarithromycin resistance.

METHODS

Three drug-resistant H. pylori strains were isolated from patients with gastritis in Bama County, China. Minimal inhibitory concentrations of clarithromycin, metronidazole, and levofloxacin were determined and complete genome sequencing was performed with annotation. Hp1181 and hp1184 genes were found in these strains and then detected by reverse transcription polymerase chain reaction. The relationships between hp1181 or hp1184 and clarithromycin resistance were ascertained with gene mutant and drug-resistant strains. The homology of the strains with hp26695 was assessed through complete genome detection and identification. Differences in genome sequences, gene quantity, and gene characteristics were detected amongst the three strains. Prediction and analysis of the function of drug-resistant genes indicated that the RNA expression of hp1181 and hp1184 increased in the three strains, which was the same in the artificially induced clarithromycin-resistant bacteria. After gene knockout, the drug sensitivity of the strains was assessed.

RESULTS

The strains showing a high degree of homology with hp26695, hp1181, and hp1184 genes were found in these strains; the expression of the genes hp1184 and hp1181 was associated with clarithromycin resistance.

CONCLUSION

Hp1181 and hp1184 mutations may be the earliest and most persistent response to clarithromycin resistance, and they may be the potential target genes for the diagnosis, prevention, and treatment of clarithromycin resistance.

Keywords: Helicobacter pylori; Clarithromycin-resistance; Diagnostic gene; Early genetic diagnosis; Helicobacter pylori strains

Core Tip: The World Health Organization designated clarithromycin-resistant Helicobacter pylori (H. pylori) a high priority among bacteria for antibiotic research and development, but the clarithromycin resistance mechanism remains unclear. We isolated and cultured clinical H. pylori strains, determined their minimal inhibitory concentrations, completed genome sequencing of hp1181 and hp1184 genes, analyzed their mutations, and found that the expression of the genes hp1184 and hp1181 was associated with clarithromycin resistance, which suggested that they can be used as genes for early diagnosis. This research may prove useful in the diagnosis, prevention, and treatment of clarithromycin-resistant H. pylori.