Colorimetric Detection of Staphylococcus aureus Based on Direct Loop-Mediated Isothermal Amplification in Combination with Lateral Flow Assay

Abstract

In this study, Staphylococcus aureus (S. aureus) was detected using a system that combined direct loop-mediated isothermal amplification (LAMP) and lateral flow assays (LFA). This technology relies on sequence-specific hybridization in LFA; furthermore, it has high specificity and addresses the limitations associated with nonspecific amplification in general colorimetric LAMP. In addition, a direct boiling method was adopted to streamline DNA extraction and enable simple detection. The established technology was used to successfully detect S. aureus at a concentration as low as 10² colony-forming unit/mL, without cross-reactivity with other strains. The practical applicability of this technology was demonstrated by analyzing real samples such as beef jerky, cabbage, and eggshell, which were artificially spiked with S. aureus. This developed system may be beneficial with regard to operational simplicity, short analysis time, and high detection performance, which would enable its application in point-of-care settings and as a novel platform for detecting various pathogens.