Rapid Detection of Streptococcus mutans Using an Integrated Microfluidic System with Loop-Mediated Isothermal Amplification

J Microbiol Biotechnol. 2023 Aug 28;33(8):1101-1110. doi: 10.4014/jmb.2304.04026. Epub 2023 May 19.

Abstract

Streptococcus mutans is the primary causative agent of caries, which is one of the most common human diseases. Thus, rapid and early detection of cariogenic bacteria is critical for its prevention. This study investigated the combination of loop-mediated isothermal amplification (LAMP) and microfluid technology to quantitatively detect S. mutans. A low-cost, rapid microfluidic chip using LAMP technology was developed to amplify and detect bacteria at 2.2-2.2 × 106 colony-forming units (CFU)/ml and its detection limits were compared to those of standard polymerase chain reaction. A visualization system was established to quantitatively determine the experimental results, and a functional relationship between the bacterial concentration and quantitative results was established. The detection limit of S. mutans using this microfluidic chip was 2.2 CFU/ml, which was lower than that of the standard approach. After quantification, the experimental results showed a good linear relationship with the concentration of S. mutans, thereby confirming the effectiveness and accuracy of the custom-made integrated LAMP microfluidic system for the detection of S. mutans. The microfluidic system described herein may represent a promising simple detection method for the specific and rapid testing of individuals at risk of caries.

Keywords: Streptococcus mutans; biosensor; dental caries; loop-mediated isothermal amplification; rapid detection.

MeSH terms

  • Humans
  • Microfluidics* / methods
  • Nucleic Acid Amplification Techniques / methods
  • Polymerase Chain Reaction
  • Streptococcus mutans* / genetics

Supplementary concepts

  • LAMP assay