Application of PCR Reverse Dot Blot Hybridization Technology for Detecting Pathogenic Bacteria in Medical Masks

To establish a rapid method utilizing PCR reverse dot blot hybridization technology for detecting pathogenic bacteria in masks. Methods: Specific genes were selected: nuc for Staphylococcus aureus, uidA for Escherichia coli, eta for Pseudomonas aeruginosa, and sly for Streptococcus hemolyticus. The bacterial 16S rDNA gene was used as a positive control. Primers and probes were designed and labeled accordingly. Probes were chemically immobilized onto a nylon membrane, and PCR products were hybridized with these fixed probes. Biotin-labeled primers bound to streptavidin, which was conjugated with horseradish peroxidase; upon reaction with a chromogenic substrate, a blue color developed. Results: The established method specifically detected pathogenic bacteria with a sensitivity ranging from 4 to 10 cfu per sample. Conclusion: The developed detection method is rapid, highly specific, and sensitive, making it suitable for the quick detection of pathogenic bacteria in masks.