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

Zhejiang Institute of Medical Device Supervision and Testing, NMPA Key Laboratory for Biomedical Optics, Key Laboratory of safety evaluation of medical devices of Zhejiang Province, Hangzhou, China

^* Corresponding author: gaonana@mdst.org.cn

Abstract

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.