MBs@SiO2-NH2-mediated magnetic separation coupled with ddPCR for rapid and simultaneous detection of four bloodstream infection pathogens

Bloodstream infections are life-threatening diseases with high morbidity and mortality. Rapid, accurate identification of causative pathogens is critical for timely targeted antimicrobial therapy. This study aimed to develop a highly sensitive analytical system for detecting major bloodstream-infection pathogens. A novel system based on amino-modified silica-coated magnetic beads was constructed to capture and concentrate Staphylococcus aureus , Enterococcus faecalis , Escherichia coli , and Klebsiella pneumoniae . Droplet digital polymerase chain reaction in a single well was applied for simultaneous absolute quantification of the four target pathogens. The proposed method completed quantitative detection of all four bacteria within 3 h. Its limit of detection reached 10 colony-forming units per milliliter in spiked blood samples. This rapid, sensitive and reliable platform enables pathogen quantification for bloodstream infections. It serves as a promising technical tool to support precise, timely diagnosis and may contribute to optimized clinical management and better patient outcomes.