Validation of loop-mediated isothermal amplification to detectHelicobacter pyloriand 23S rRNA mutations: A prospective, observational clinical cohort study

Background Identifying point mutations in 23S rRNA closely associated with clarithromycin resistance can increase the eradication rate ofHelicobacter pylori(H pylori). In this study, we verified the sensitivity, specificity, and reliability of a newly developed loop-mediated isothermal amplification (LAMP) assay kit to detectH pyloriand 2143G and 2182C mutations in 23S rRNA. Methods LAMP assay to detectH pyloriand a mutant strain with 2143G and 2182C was conducted with the Isopollo(R)H pylori& ClaR kit. A prospective, open-label, observational study was conducted to validate the reliability of the LAMP assay in both a development cohort and a bedside direct LAMP cohort. Results The LAMP assay had good sensitivity, as it could detect as few as 10-100 copies ofH pyloriand mutants with 2143G and 2182C in 23S rRNA, and good specificity, as it did not react with other bacterial species. In the development cohort with 622 participants, the LAMP assay showed good agreement with RUT for detectingH pylori(kappa value 0.923,P < .001) and had exactly the same results as sequencing analysis for 2143G and 2182C point mutations. The direct LAMP cohort including 93 patients had 97.7% (42/43) of concordance in detecting 2143G and 2182C point mutations compared to the PCR-based sequencing analysis. Conclusion The Isopollo(R)H pylori& ClaR LAMP assay was a valid method for detectingH pyloriand for 2143G and 2182C point mutations in 23S rRNA in a clinical setting.

Automated summary

The newly developed LAMP assay kit demonstrated good sensitivity, specificity, and reliability in detecting H pylori and 2143G and 2182C mutations in 23S rRNA. The study showed high agreement with traditional detection methods and sequencing analysis, making it a valid method for clinical detection of H pylori and point mutations.