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Gastric Mucosa Quantitative Polymerase Chain Reaction Analysis for Detecting Helicobacter pylori and Antibiotic Resistance
Published on: March 7, 2025
Early genetic diagnosis of clarithromycin resistance in Helicobacter pylori
Xiao-Hua Li1, Yong-Yi Huang1, Lin-Ming Lu2
1Research Center for the Prevention and Treatment of Drug Resistant Microbial Infection, Youjiang Medical University for Nationalities, Baise 533000, Guangxi Zhuang Autonomous Region, China.
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.
Insights
Drug resistance in Helicobacter pylori (H. pylori) is rising. This study identifies Hp1181 and Hp1184 gene mutations as potential early indicators for clarithromycin resistance, aiding in diagnosis and treatment strategies.
Area of Science:
- Microbiology
- Genomics
- Drug Resistance Research
Background:
- Increasing rates of drug resistance in clinical Helicobacter pylori (H. pylori) isolates pose a significant challenge.
- The precise mechanisms underlying H. pylori drug resistance remain largely unelucidated.
- This study focused on genomic analysis of drug-resistant H. pylori strains from diverse Chinese populations.
Purpose of the Study:
- To identify specific drug-resistant genes within H. pylori.
- To pinpoint genes crucial for the early diagnosis of clarithromycin resistance.
- To investigate the role of Hp1181 and Hp1184 in H. pylori clarithromycin resistance.
Main Methods:
- Isolation and whole-genome sequencing of three drug-resistant H. pylori strains.
- Determination of minimal inhibitory concentrations for key antibiotics (clarithromycin, metronidazole, levofloxacin).
- Reverse transcription polymerase chain reaction (RT-PCR) to detect Hp1181 and Hp1184 gene expression, alongside gene knockout experiments to assess drug sensitivity.
Main Results:
- High genomic homology was observed between the isolated strains and the reference strain hp26695.
- The expression levels of Hp1181 and Hp1184 genes were found to correlate significantly with clarithromycin resistance.
- Increased RNA expression of Hp1181 and Hp1184 was noted in resistant strains, mirroring artificially induced resistance.
Conclusions:
- Mutations in Hp1181 and Hp1184 represent potential early and persistent markers of clarithromycin resistance in H. pylori.
- These genes are identified as promising targets for the development of diagnostic tools for clarithromycin resistance.
- Targeting Hp1181 and Hp1184 could facilitate improved prevention and treatment strategies for H. pylori infections.
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