A novel loop-mediated isothermal amplification-lateral flow dipstick method for Helicobacter pylori detection

Area of Science:

• Microbiology
• Molecular Diagnostics
• Biotechnology

Background:

• Helicobacter pylori (H. pylori) infection is a major risk factor for gastric cancer.
• Current detection methods for H. pylori lack sensitivity, specificity, or convenience.
• A simple, rapid, and accurate diagnostic tool is needed for H. pylori eradication.

Purpose of the Study:

• To develop and validate a novel loop-mediated isothermal amplification-lateral flow dipstick (LAMP-LFD) assay for H. pylori detection.
• To assess the sensitivity, specificity, and clinical applicability of the LAMP-LFD method.

Main Methods:

• Primers targeting the conserved ureB gene of H. pylori were designed for LAMP.
• Biotin-labeled UreB-FIP was used for amplification, and FAM-labeled probes were detected by LFD.
• A clinical study evaluated the LAMP-LFD assay using 20 fecal samples.

Main Results:

• The optimized LAMP-LFD assay detected H. pylori specifically at 65°C for 60 min, with no cross-reactivity.
• The assay demonstrated a lower limit of detection of 10^2 copies/μL, 100 times more sensitive than PCR.
• H. pylori was detected in 13 out of 20 fecal samples using the LAMP-LFD method.

Conclusions:

• A novel LAMP-LFD assay for H. pylori detection has been successfully established.
• The assay is highly sensitive, specific, and operates simply within 1.5 hours.
• This method presents a promising tool for H. pylori diagnosis in primary healthcare and low-resource settings.