Detection of ESKAPE Bacterial Pathogens at the Point of Care Using Isothermal DNA-Based Assays in a Portable Degas-Actuated Microfluidic Diagnostic Assay Platform
Abstract
An estimated 1.5 billion microbial infections occur globally each year and result in ∼4.6 million deaths. A technology gap associated with commercially available diagnostic tests in remote and underdeveloped regions prevents timely pathogen identification for effective antibiotic chemotherapies for infected patients. The result is a trial-and-error approach that is limited in effectiveness, increases risk for patients while contributing to antimicrobial drug resistance, and reduces the lifetime of antibiotics. This paper addresses this important diagnostic technology gap by describing a low-cost, portable, rapid, and easy-to-use microfluidic cartridge-based system for detecting the ESKAPE ( Enterococcus faecium , Staphylococcus aureus , Klebsiella pneumoniae , Acinetobacter baumannii , Pseudomonas aeruginosa , and Enterobacter spp.) bacterial pathogens that are most commonly associated with antibiotic resistance. The point-of-care molecular diagnostic system consists of a vacuum-degassed microfluidic cartridge preloaded with lyophilized recombinase polymerase amplification (RPA) assays and a small portable battery-powered electronic incubator/reader. The isothermal RPA assays detect the targeted ESKAPE pathogens with high sensitivity (e.g., a limit of detection of ∼10 nucleic acid molecules) that is comparable to that of current PCR-based assays, and they offer advantages in power consumption, engineering, and robustness, which are three critical elements required for the point-of-care setting.
Document Details
- Document Type
- Pub Defense Publication
- Publication Date
- Feb 15, 2017
- Source ID
- 10.1128/aem.02449-16
Entities
People
- Adam C. Siegel
- Clint Torres
- Douglas B. Weibel
- Jindong Zan
- Lars David Renner
- Linda I. Hu
- Manuel Martínez
- Pedro J. Resto
- Sara B. Hall
- Tom R. Slezak
- Tuan H. Nguyen
Organizations
- Gates Foundation
- Lawrence Livermore National Laboratory
- Leibniz Institute for Polymer Research
- National Science Foundation
- United States Department of Defense
- University of Wisconsin–Madison