Rapid Detection of Bacterial Pathogens Causing Lower Respiratory Tract Infections via Microfluidic-Chip-Based Loop-Mediated Isothermal Amplification

J Vis Exp. 2024 Mar 29:(205). doi: 10.3791/66677.

Abstract

Respiratory tract infections (RTIs) are among the most common problems in clinical settings. Rapid and accurate identification of bacterial pathogens will provide practical guidelines for managing and treating RTIs. This study describes a method for rapidly detecting bacterial pathogens that cause respiratory tract infections via multi-channel loop-mediated isothermal amplification (LAMP). LAMP is a sensitive and specific diagnostic tool that rapidly detects bacterial nucleic acids with high accuracy and reliability. The proposed method offers a significant advantage over traditional bacterial culturing methods, which are time-consuming and often require greater sensitivity for detecting low levels of bacterial nucleic acids. This article presents representative results of K. pneumoniae infection and its multiple co-infections using LAMP to detect samples (sputum, bronchial lavage fluid, and alveolar lavage fluid) from the lower respiratory tract. In summary, the multi-channel LAMP method provides a rapid and efficient means of identifying single and multiple bacterial pathogens in clinical samples, which can help prevent the spread of bacterial pathogens and aid in the appropriate treatment of RTIs.

Publication types

  • Video-Audio Media

MeSH terms

  • Humans
  • Klebsiella pneumoniae
  • Microfluidics
  • Molecular Diagnostic Techniques*
  • Nucleic Acid Amplification Techniques*
  • Nucleic Acids*
  • Reproducibility of Results
  • Respiratory Tract Infections* / diagnosis

Substances

  • Nucleic Acids

Supplementary concepts

  • LAMP assay