Ratiometric Multiplexed PCR Assay on a Portable Platform for Bacterial Identification from Urine

Abstract

We present a fully integrated, portable, and multiplexed real-time PCR droplet magnetofluidic platform capable of identifying up to 6 bacterial species directly from urine samples. Rapid identification of pathogens at the point-of-care (POC) with our platform can inform targeted treatments to combat the emergence of antibiotic resistant pathogens. Our platform provides a solution to the need of multiplexed detection capacity for diagnosis of infectious diseases, especially in which a broad range of bacteria may be present. The multiplexed capacity is achieved by a ratiometric real-time PCR assay that contains universal primers and species-specific TaqMan probes targeting variable region of 16S ribosomal RNA of each bacterium. The detection capacity is demonstrated with the analysis of six common bacteria in urogenital infections with input concentrations from 104 to 107 copies/mL to show the potential diagnostic value.