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

Yi Zhu,1,2 Chunhui Lin,1,2 Huaming Xu,3 Zhaoxin Xia,1,2 Wensu Yang,1,2 Hao Tang,4 Xinyi Hu,1,2 Tong Jiang,1,2 Zhen Liu,1,2 Jilu Shen1,2 1The First Affiliated Hospital of Anhui Medical University, Hefei, People’s Republic of China; 2Anhui Public Health Clinical Center, Hefei, People’s Republic of China; 3The First Affiliated Hospital of Anhui University of Chinese Medicine, Hefei, People’s Republic of China; 4The Second Affiliated Hospital of Anhui Medical University, Hefei, People’s Republic of ChinaCorrespondence: Jilu Shen, Tel +86 151 5515 2963, Email shenjilu@ahmu.edu.cn; 1464675852@qq.comIntroduction: 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