Establishment and Methodological Evaluation of a Method for Rapid Detection of Helicobacter pylori and Virulence Genes Based on CRISPR-Cas12a,Infection and Drug Resistance

Introduction: More than half of the world’s people are infected or have been infected with Helicobacter pylori. This infection is related to many diseases, with its pathogenicity related to virulence factors. Therefore, the rapid diagnosis of H. pylori and genotyping of virulence genes play an extremely important role in the clinical treatment and control of transmission.
Methods: To this end, we developed a molecular detection method based on RPA- CRISPR-Cas12a technology for the specific genes 16S rDNA gene, cytotoxin associated gene A(cagA), and vacuolating cytotoxin A (vacA) of H. pylori.
Results: The results of which were displayed by lateral flow strips. Macroscopic observation takes only about 25 minutes and the sensitivity is 2ng/microliter.
Discussion: The method is simple, convenient to operate and has low costs, and can therefore be applied widely to the detection and typing of H. pylori in various environments such as primary hospitals, community clinics, outdoors, and large medical institutions.

Keywords: Helicobacter pylori, virulence genes, recombinase polymerase amplification, CRISPR-Cas12a, lateral flow immunochromatographic strip

中文翻译:








基于CRISPR-Cas12a的幽门螺杆菌及毒力基因快速检测方法的建立及方法学评价



简介:世界上超过一半的人感染或曾经感染过幽门螺杆菌。这种感染与多种疾病有关,其致病性与毒力因子有关。因此,H. pylori的快速诊断和毒力基因分型在临床治疗和传播控制中具有极其重要的作用。
方法:为此,我们开发了一种基于RPA-CRISPR-Cas12a技术的幽门螺杆菌特异性基因16S rDNA基因、细胞毒素相关基因A(cagA)和空泡细胞毒素A(vacA)的分子检测方法
结果:其结果由侧流条显示。宏观观察仅需25分钟左右,灵敏度为2ng/微升。
讨论:该方法简单、操作方便、成本低,可广泛应用于基层医院、社区门诊、户外、大型医疗机构等各种环境的幽门螺杆菌检测分型。

关键词: 幽门螺杆菌,毒力基因,重组酶聚合酶扩增,CRISPR-Cas12a,侧流免疫层析试纸条






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