Integrated Bacterial Identification and Antimicrobial Susceptibility Testing for Polymicrobial Infections Using Digital PCR and Digital High-Resolution Melt in a Microfluidic Array Platform.

In diagnosing bacterial infection, rapid bacterial identification (ID) and antimicrobial susceptibility testing (AST) are critical to clinicians in order to provide an effective treatment in a timely manner. The gold standard, culture-based approach provides both ID and antimicrobial susceptibility but requires several days of turnaround time. Especially in polymicrobial infections, where there are more than one organisms interacting collectively that can complicate the treatment. Here, we demonstrate a rapid bacterial diagnostic approach that is capable of bacterial ID/AST in heterogeneous samples within less than 4 hours by using digital PCR (dPCR) and digital high-resolution melt via microfluidic devices. By utilizing dPCR, we are able to quantify amount of nucleic acid, which correlates to phenotypic responses of {\bf individual pathogens in a mixed sample and also shorten the required time of antibiotic exposure. In addition, we employ a machine learning algorithm to automatically identify bacterial species based on melt profiles of 16S rRNA gene.