Efficient Detection Of S. Aureus Using Lamp-Pfago System

Oral and maxillofacial infections are diseases resulting from the invasion and colonization of pathogens in the oral and maxillofacial tissues. These infections elicit an inflammatory response predominantly driven by the host's defense mechanisms, ultimately leading to damage of the oral tissues. Notably, these infections are characterized by their sudden onset and rapid progression. One of the most common pathogenic bacteria that cause infection is Staphylococcus aureus. Therefore, in this study, the method combining the LAMP with Pyrococcus furiosus Argonaute (PfAgo) in a one tube form was utilized to rapidly, sensitively and specifically detect S. aureus, and to investigate its detection performance. In a one tube, the specific nuc gene fragment of S. aureus was first amplified using LAMP. Subsequently, the amplified fragment was combined with PfAgo, and the PfAgo activity was stimulated to cleave the fluorescent probe and emit fluorescence, thereby completing the detection of the target gene. The one-tube LAMP-PfAgo system is capable of completing the detection within 48 minutes, and this system can detect bacterial solutions within the concentration range of 10⋀0CFU/mL to 10⋀7CFU/mL, and a limit of detection (LOD) of 10⋀0CFU/mL. Moreover, the concentration of the bacterial solution shows a good linear relationship with the fluorescence intensity. Meanwhile, five different bacteria were selected for specific detection, and the results indicated that this system has a good specificity recognition ability. The one-tube LAMP-PfAgo system offers an efficient, specific, and portable innovative approach for detecting S. aureus.