Vancomycin-dendrimer based multivalent magnetic separation nanoplatforms combined with multiplex quantitative PCR assay for detecting pathogenic bacteria in human blood

Sepsis caused by bacteria has high morbidity and mortality, and it is neccerssay to establish a fast, convenient, and facility assays for detection of bacteria. In this study, we have developed established a simple, rapid, and ultrasensitive vancomycin (Van) and dendrimer nanoparticles-based method to isolate and detect bacteria in human blood using a multivalent binding strategy. The proposed Bio-den-Van multivalent capture nanoplatform combined with m-qPCR for simultaneous detection of two kinds of bacteria was demonstrated with rapid 2 min bacteria isolation with a linear range at 3.2 × 10¹-3.2 × 10⁶ CFU·mL^-1 for L. monocytogenes and 4.1 × 10¹-4.1 × 10⁶ CFU·mL^-1 for S. aureus, respectively. The limit of detection (LOD) for simultaneous detection of L. monocytogenes and S. aureus were 32 and 41 CFU·mL^-1 in spiked human blood samples, respectively. Other bacteria had an insignificant interference with the test results. This Bio-den-Van multivalent capture nanoplatform combined with m-qPCR detection exhibited rapid, high sensitivity and specificity in simultaneous detection of various bacteria. To our knowledge, this is the first time that Bio-den-Van multivalent capture nanoplatform was used with Van as a recognition molecule for the simultaneous capture and subsequent detection of two bacteria from spiked human blood sample. This method holds great potential for future clinical applications.