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

幽门螺杆菌 聚合酶链反应 微生物学 抗生素耐药性 抗生素 螺杆菌感染 生物 基因 遗传学
作者
Jinxin Lai,Huimin Chen,Cheng-Yuan He,Yu‐Ting Si,Luan Luan,Ge Huang,You‐Wei Zheng,Yuwei Di,Xinqiang Zhang,Yan‐an Yao,Zheng‐Kang Li,Liang Wang
出处
期刊:Journal of Visualized Experiments [MyJOVE]
卷期号: (219)
标识
DOI:10.3791/67710
摘要

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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