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 min 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 min with a limit of detection of 1000 bacteria in 1 mL urine.
Bibliographical noteFunding Information:
The authors would like to thank the anonymous volunteers who provided urine samples for the duration of the study. This work has been supported by National Institute for Health Research (NIHR), Rapid detection of infectious agents at point of triage (PoT), II-ES-0511-21002. The views expressed in this publication are those of the authors and not necessarily those of the NHS, the National Institute for Health Research, Public Health England or the Department of Health.
© 2016, The Author(s).
- Antibiotic resistant bacteria
- Magnetic beads
- Recombinase polymerase amplification
- Sample preparation