4.4 Article

Non-Invasive Colorimetric Magneto Loop-Mediated Isothermal Amplification (CM-LAMP) Method for Helicobacter pylori Detection

Journal

JOURNAL OF MICROBIOLOGY AND BIOTECHNOLOGY
Volume 31, Issue 4, Pages 501-509

Publisher

KOREAN SOC MICROBIOLOGY & BIOTECHNOLOGY
DOI: 10.4014/jmb.2101.01008

Keywords

Loop-mediated isothermal amplification (LAMP); colorimetric magneto LAMP assay (CM-LAMP); magnetic nanoparticles (MNPs); Helicobacter pylori; non-invasive test; saliva

Funding

  1. National Research Council of Thailand (NRCT) through the Royal Golden Jubilee PhD Program [PHD/0224/2558]
  2. Thailand Science Research and Innovation (TSRI) through the Royal Golden Jubilee PhD Program [PHD/0224/2558]
  3. Thailand Research Fund (TRF) through the Research Grant for New Scholar [MRG5580110]
  4. Prince of Songkla University [MET6302042S]

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More than half the world's population is thought to be infected with Helicobacter pylori, which can cause gastric ulcers and deadly gastric cancer. This study presents a highly specific and sensitive noninvasive assay for detecting H. pylori in saliva samples, offering a potential alternative to current invasive diagnostic methods.
More than half the world's population is thought to be infected with Helicobacter pylori. Although the majority of infected people are asymptomatic, H. pylori infection may cause gastric ulcers and deadly gastric cancer. Owing to the difficulty and invasiveness of current routine culture and diagnostic methods, a highly sensitive and specific noninvasive assay for H. pylori is of interest. This study highlighted the design and performance of a colorimetric magneto loop-mediated isothermal amplification (CM-LAMP) assay to detect H. pylori in spiked saliva samples. LF primers were coated on magnetic nanoparticles by carbodiimide-induced immobilization and functionally used for solid phase amplification. During the LAMP reaction at 66 degrees C, biotin-tagged FIPs were incorporated into LAMP amplicons. The colorimetric signal developed after the addition of NeutrAvidin horseradish peroxidase conjugate (NA-HRP) and ABTS. None of the tested microorganisms, including closely related bacteria, was shown positive by the CM-LAMP assay except H. pylori isolates. This novel platform was highly specific and 100-fold more sensitive (40 CFU/ml or 0.2 CFU per reaction) than the PCR and conventional LAMP assays for the detection of H. pylori in spiked saliva. Our results demonstrated the feasibility of using this noninvasive molecular diagnostic test to detect H. pylori in saliva samples.

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