Simultaneous Detection of Helicobacter pylori and Clarithromycin Resistance Mutations Using RAA-CRISPR/Cas13a Assay

被引：4

作者：

Yan, Kuocheng ^{[1
]}

Wang, Xiaohui ^{[2
]}

Han, Yao ^{[1
]}

Tian, Yuan ^{[1
]}

Niu, Mengwei ^{[1
]}

Dong, Xue ^{[1
]}

Li, Xiaowei ^{[2
]}

Li, Hao ^{[1
]}

Sun, Yansong ^{[1
]}

机构：

[1] Beijing Inst Microbiol & Epidemiol, State Key Lab Pathogen & Biosecur, Beijing 100071, Peoples R China

[2] Chinese Peoples Liberat Army Gen Hosp, Med Ctr 6, Dept Gastroenterol, Beijing 100080, Peoples R China

来源：

INFECTION AND DRUG RESISTANCE | 2024年 / 17卷

关键词：

Helicobacter pylori; clarithromycin resistance mutation; recombinase aided amplification; CRISPR/Cas13a; nucleic acid; detection; PCR;

D O I：

10.2147/IDR.S462963

中图分类号：

R51 [传染病];

学科分类号：

100401 ;

摘要：

Background: Infection caused by Helicobacter pylori (H. pylori) affects approximately 50% of the global population. It is a major pathogenic factor for chronic gastritis and gastric cancer. Besides, the resistance to antibiotics such as clarithromycin could reduce the eradication rate. Currently, there is an urgent need for a swift, easy to perform, and highly sensitive detection method for H. pylori and clarithromycin resistance. Methods: We used FAM/Digoxin labeled primers to amplify specific H. pylori 23S rRNA fragments by Recombinase Aided Amplification (RAA), and resistance mutations were distinguished using CRISPR/Cas13a system combined with lateral flow strip. Twenty-eight saliva samples were analyzed using qPCR, gene sequencing and this method to evaluate the detection efficiency. Results: We developed a simultaneous detection method for H. pylori and clarithromycin resistance mutations named sensitive H. pylori easy-read dual detection (SHIELD). The results showed both A2142G and A2143G mutant DNAs causing clarithromycin resistance could be distinguished from the wild type with a concentration of 50 copies/mu L, and no cross-reaction with other 5 common gastrointestinal bacteria was observed. For the detection of H. pylori in 28 saliva samples, the positive predictive value of this method was 100% (19/19) in comparison with qPCR. For detecting clarithromycin resistance, the positive predictive value of this method was 84.6% (11/13) compared with gene sequencing. Conclusion: SHIELD assay showed high sensitivity and specificity in detecting H. pylori and clarithromycin resistance mutations. It could be a potential measure in the rapid detection of H. pylori, large-scale screening and guiding clinical medication.

引用

页码：3001 / 3010

页数：10

共 29 条

[1] Diagnosis of Helicobacter pylori Using Invasive and Noninvasive Approaches [J].