Portable loop-mediated isothermal amplification device with spectrometric detection for rapid pathogen identification

Anal Biochem. 2024 Nov:694:115615. doi: 10.1016/j.ab.2024.115615. Epub 2024 Jul 11.

Abstract

With the rise in extreme weather due to global warming, coupled with globalization facilitating the spread of infectious diseases, there's a pressing need for portable testing platforms offering simplicity, low cost, and remote transmission, particularly beneficial in resource-limited and non-urban areas. We have developed a portable device using loop-mediated isothermal amplification (LAMP) with spectrometric detection to identify Salmonella Typhimurium DNA. The device utilizes the LinkIt 7697 microcontroller and a microspectrometer to capture and transmit spectral signals in real-time, allowing for improved monitoring and analysis of the reaction progress. We built a hand-held box containing a microspectrometer, thermoelectric cooler, ultraviolet LED, disposable reaction tube, and homemade thermal module, all powered by rechargeable batteries. Additionally, we conducted thorough experiments to ensure temperature accuracy within 1 °C under thermal control, developed a heating module with a LinkIt 7697 IoT development board to heat the DNA mixture to the reaction temperature within 3 min, and integrated foam insulation and a 3D-printed frame to enhance the device's thermal stability. We successfully demonstrated the amplification of Salmonella Typhimurium DNA with an impressive sensitivity of 2.83 × 10-4 ng/μL. A remote webpage interface allows for monitoring the temperature and fluorescence during the LAMP process, improving usability. This portable LAMP device with real-time detection offers a cost-effective solution for detecting Salmonella Typhimurium in food products. Its unique design and capabilities make it a promising tool for ensuring food safety.

Keywords: Infectious diseases; Loop-mediated isothermal amplification (LAMP); Portable testing platform; Salmonella typhimurium DNA; Spectrometric detection.

MeSH terms

  • DNA, Bacterial* / analysis
  • DNA, Bacterial* / genetics
  • Food Microbiology
  • Molecular Diagnostic Techniques
  • Nucleic Acid Amplification Techniques* / instrumentation
  • Nucleic Acid Amplification Techniques* / methods
  • Salmonella typhimurium* / genetics
  • Salmonella typhimurium* / isolation & purification

Substances

  • DNA, Bacterial

Supplementary concepts

  • LAMP assay