Simple and Rapid Sample Preparation System for the Molecular Detection of Antibiotic Resistant Pathogens in Human Urine

Overview

Journal Biomed Microdevices

Specialty Biomedical Engineering

Date 2016 Feb 6

PMID 26846875

Citations 4

Authors

Martha Valiadi

Sumit Kalsi

Isaac G F Jones

Carrie Turner

J Mark Sutton

Hywel Morgan

Affiliations

Soon will be listed here.

Abstract

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 (bla CTX-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 bla CTX-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.

Citing Articles

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Development of a panel of recombinase polymerase amplification assays for detection of common bacterial urinary tract infection pathogens.

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