• Developed TPMSR, a triple-plasmid-mediated synchronous recombination strategy that enhances phage genome editing efficiency by optimizing homologous arm length and cleavage pressure, overcoming limitations of high-activity sgRNA dependence.
• Engineered reporter phage T7::Nluc by integrating the Nluc gene into phage T7, enabling sensitive detection of E. coli in blood samples.
• Integrated T7::Nluc into a microfluidic chip platform, achieving rapid detection of E. coli in blood within 1.5 hours at concentrations below 30 CFU/mL.
• Validated the platform with 51 clinical isolates, demonstrating outstanding sensitivity, specificity, and accuracy, offering a promising tool for rapid diagnosis of bloodstream infections.