The Accuracy of Melting Curve–Based Multiplex Real-Time PCR for Diagnosing Helicobacter pylori Resistance to Clarithromycin and Levofloxacin in Stool Specimens

IF 4.3 2区医学 Q1 GASTROENTEROLOGY & HEPATOLOGY

Helicobacter Pub Date : 2025-02-11 DOI:10.1111/hel.70012

Youhua Wang, Xiaoling Gong, Qirui Lai, Hui Fang, Xiyin Yang, Xuan Li, Dong Sheng Liu, Yong Xie

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引用次数: 0

Abstract

Aims

This study evaluates the accuracy of melting curve–based multiplex real-time PCR (multiplex rt-PCR) on stool samples for diagnosing antibiotic resistance in Helicobacter pylori (H. pylori) compared to E-test and sequencing.

Methods

Gastric biopsies and stool samples were collected from 385 H. pylori-infected patients. A total of 325 strains were isolated, and genomic DNA was extracted from all 385 stool samples. E-tests were conducted to detect phenotypic resistance for clarithromycin and levofloxacin. Sanger sequencing and multiplex rt-PCR were employed to identify H. pylori 23S rRNA and GyrA mutations.

Results

E-test results indicated that 203 (62.5%) were susceptible to both clarithromycin and levofloxacin, 33 (10.2%) exhibited mono-resistance to clarithromycin, 48 (14.8%) showed mono-resistance to levofloxacin, and 41 (12.6%) had dual resistance to both antibiotics. Compared to E-test results, the sensitivity and specificity of the multiplex rt-PCR method for detecting clarithromycin resistance mutation were 93.2 (95% CI 84.3–97.5) and 87.1% (95% CI 82.2–90.9), respectively. For levofloxacin resistance mutation, the multiplex rt-PCR method showed a sensitivity of 80.7 (95% CI 70.3–88.3) and a specificity of 93.0% (95% CI 88.7–95.8). Compared to Sanger sequencing, the sensitivity and specificity of the multiplex rt-PCR method for detecting clarithromycin resistance mutation were 95.8 (95% CI 90.0–98.4) and 96.0% (95% CI 92.6–98.0), respectively. For levofloxacin resistance mutation, the multiplex rt-PCR method showed a sensitivity of 91.3% (95% CI, 83.1–95.9) and a specificity of 96.1% (95% CI, 92.7–98.0).

Conclusion

Genotypic methods, including Sanger sequencing and multiplex rt-PCR, were rapid and reliable for diagnosing clarithromycin and levofloxacin resistance in the stool samples.

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来源期刊

Helicobacter 医学-微生物学

CiteScore

8.40

自引率

9.10%

发文量

审稿时长

2 months

期刊介绍： Helicobacter is edited by Professor David Y Graham. The editorial and peer review process is an independent process. Whenever there is a conflict of interest, the editor and editorial board will declare their interests and affiliations. Helicobacter recognises the critical role that has been established for Helicobacter pylori in peptic ulcer, gastric adenocarcinoma, and primary gastric lymphoma. As new helicobacter species are now regularly being discovered, Helicobacter covers the entire range of helicobacter research, increasing communication among the fields of gastroenterology; microbiology; vaccine development; laboratory animal science.