Vancomycin-dendrimer based multivalent magnetic separation nanoplatforms combined with multiplex quantitative PCR...

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.