Detection of <em>Helicobacter pylori</em> Infection and Antibiotic Resistance via Stool Quantitative Polymerase Chain Reaction Analysis

Helicobacter pylori (H. pylori) is widely prevalent worldwide, with approximately 50% of the global population having a history of H. pylori infection. In China, the infection rate ranges from 40% to 70%. H. pylori is primarily associated with gastrointestinal diseases such as chronic gastritis, gastric ulcers, and duodenal ulcers. Currently, the clinical treatment for H. pylori infection involves either triple or quadruple therapy. However, the extensive use of antibiotics has led to the development of antibiotic resistance in H. pylori. Therefore, detecting both H. pylori and its antibiotic resistance is crucial for guiding clinical treatment. Diagnostic methods for H. pylori include urea breath test (UBT), antigen test, serological antibody test, endoscopy, rapid urease test (RUT), and bacterial culture. While the first three methods are non-invasive, they do not allow for bacterial recovery and, thus, cannot be used for resistance testing. The latter three methods are invasive, expensive, require high technical expertise, and may cause harm to patients. Therefore, a non-invasive, rapid method for simultaneous detection of H. pylori infection and antibiotic resistance is of utmost clinical importance for the effective eradication of H. pylori. This paper aims to introduce a specific protocol that combines quantitative polymerase chain reaction (qPCR) with TaqMan fluorescent probe technology to rapidly detect H. pylori infection and antibiotic resistance. This method provides a convenient, rapid, and non-invasive way to diagnose H. pylori infection and resistance, unlike traditional bacterial culture and other techniques. qPCR is used to identify the infection and detect mutations in the 23S rRNA and gyrA genes, which are linked to resistance to clarithromycin and quinolones, respectively. Compared to conventional culture techniques, this approach offers a non-invasive, cost-effective, and time-efficient method for detecting Helicobacter pylori infection and determining its antibiotic resistance.

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Publication Details

Journal
Journal of Visualized Experiments
Published
2025-05-16
DOI
https://doi.org/10.3791/67710
Primary Topic
Helicobacter pylori-related gastroenterology studies
Type
article
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article

Detection of Helicobacter pylori Infection and Antibiotic Resistance via Stool Quantitative Polymerase Chain Reaction Analysis

Xinqiang Zhang, Ge Huang, Yan‐an Yao, Yuwei Di et al.
Journal of Visualized Experiments
Helicobacter pylori-related gastroenterology studies
article

Detection of Helicobacter pylori Infection and Antibiotic Resistance via Stool Quantitative Polymerase Chain Reaction Analysis

Xinqiang Zhang, Ge Huang, Yan‐an Yao, Yuwei Di, Yu‐Ting Si, Cheng-Yuan He, Jinxin Lai, Luan Luan, Zheng‐Kang Li, Huimin Chen, Liang Wang, You‐Wei Zheng
article en

Abstract

Helicobacter pylori (H. pylori) is widely prevalent worldwide, with approximately 50% of the global population having a history of H. pylori infection. In China, the infection rate ranges from 40% to 70%. H. pylori is primarily associated with gastrointestinal diseases such as chronic gastritis, gastric ulcers, and duodenal ulcers. Currently, the clinical treatment for H. pylori infection involves either triple or quadruple therapy. However, the extensive use of antibiotics has led to the development of antibiotic resistance in H. pylori. Therefore, detecting both H. pylori and its antibiotic resistance is crucial for guiding clinical treatment. Diagnostic methods for H. pylori include urea breath test (UBT), antigen test, serological antibody test, endoscopy, rapid urease test (RUT), and bacterial culture. While the first three methods are non-invasive, they do not allow for bacterial recovery and, thus, cannot be used for resistance testing. The latter three methods are invasive, expensive, require high technical expertise, and may cause harm to patients. Therefore, a non-invasive, rapid method for simultaneous detection of H. pylori infection and antibiotic resistance is of utmost clinical importance for the effective eradication of H. pylori. This paper aims to introduce a specific protocol that combines quantitative polymerase chain reaction (qPCR) with TaqMan fluorescent probe technology to rapidly detect H. pylori infection and antibiotic resistance. This method provides a convenient, rapid, and non-invasive way to diagnose H. pylori infection and resistance, unlike traditional bacterial culture and other techniques. qPCR is used to identify the infection and detect mutations in the 23S rRNA and gyrA genes, which are linked to resistance to clarithromycin and quinolones, respectively. Compared to conventional culture techniques, this approach offers a non-invasive, cost-effective, and time-efficient method for detecting Helicobacter pylori infection and determining its antibiotic resistance.

Journal of Visualized Experiments(219)
Xuzhou Medical College (CN), Guangdong Academy of Medical Sciences (CN), Guangdong Provincial People's Hospital (CN), Southern Medical University (CN)
Openalex Percentile: Top 81%
Helicobacter pylori-related gastroenterology studies
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