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

Abstract

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

Ethics Approval and Consent to Participate

This study involved the use of the patient’s saliva, which had been given with the informed consent of the subjects, and all specimens were numerically numbered and did not affect the patient’s privacy. All procedures performed in studies involving human participants were in accordance with the ethical standards of the medical ethics committee of the First Affiliated Hospital of Anhui Medical University (Anhui Public Health Clinical Center)(reference number: LISC20210802) and with the 1964 Helsinki Declaration and its later amendments or comparable ethical standards.

Acknowledgments

The authors would like to express their gratitude to EditSprings for the expert linguistic services provided.

Author Contributions

All authors made a significant contribution to the work reported, whether that is in the conception, study design, execution, acquisition of data, analysis and interpretation, or in all these areas; took part in drafting, revising, or critically reviewing the article; gave final approval of the version to be published; have agreed on the journal to which the article has been submitted; and agree to be accountable for all aspects of the work.

Disclosure

The authors declare that they have no competing interests for this work.

Additional information

Funding

This work was funded through a grant from Anhui Provincial Department of Education for university cooperative research and public health collaborative innovation project in Anhui Province in 2020 (Grant No. GXXT-2020-016) and a grant from Anhui Provincial Health Commission for key scientific research projects in 2021 (Grant No. AHWJ2021a011).