Simple and rapid sample preparation system for the molecular detection of antibiotic resistant bacteria in human urine

Antibiotic resistance in urinary tract infections (UTIs) can cause significant complications without quick detection and appropriate treatment. . We describe a new approach to capture, concentrate and prepare amplification-ready DNA from antibiotic resistant bacteria in human urine samples. Klebsiella pneumoniae NCTC13443 (blaCTX-M-15 positive) spiked into filtered human urine was used as a model system. Bacteria were captured using anion exchange diaethylaminoethyl (DEAE) magnetic microparticles and concentrated 200-fold within ~3.5 minutes using a custom, valve-less microfluidic chip. Eight samples were processed in parallel, and DNA, was released using heat lysis from an integrated resistive heater. The crude cell lysate was used for real time Recombinase Polymerase Amplification (RPA) of the blaCTX-M-15 gene. The end to end processing time was approximately 15 minutes with a limit of detection of 1000 bacteria in 1 mL urin