Background

A large percentage of the 8 million urinary tract infections (UTI’s) that occur each year in the U.S. are caused by E. coli. Increasingly, these bacteria-driven UTI’s are resistant to available antibiotics and lead to severe complications. Some studies have indicated that patients could be contracting UTI’s through consumption of tainted meat and produce http://www.pbs.org/wgbh/frontline/article/can-e-coli-in-supermarket-meat-cause-utis/; http://news.ubc.ca/2015/02/04/antibiotic-resistant-e-coli-detected-on-vancouver-farmers-market-produce/).

The 2014 WHO release Antimicrobial Resistance: Global Report on Surveillance observed that fluoroquinolones were ineffective in more than half of patients with UTI’s caused by E. coli. Early and specific identification of drug resistant E. coli is thus critical in formulating effective patient treatment plans.

Patient treatments can be delayed due to lengthy pathogen identification techniques. Detection of a suspected pathogen is currently performed using urine cultures microscopy, leukocyte esterase and tests. These tests can take 1-2 days to complete and require microbiological lab facilities. Simple (CLIA-waved) molecular diagnostics for bedside testing can significantly reduce time to treatment.

Description

LLNL researchers are developing a battery-powered molecular diagnostic (MDx) platform for biothreat and public health agent identification. LLNL researchers developed isothermal molecular diagnostic assays to detect and identify DNA from pathogenic bacteria, including E. coli, known to cause UTI’s and sepsis. Detection of the bacterial DNA does not require technically challenging DNA isolation and purification.

Additional technical information can be found in the Journal of Microbiological Methods published article, Detection of Bacillus anthracis from spores and cells by loop-mediated isothermal amplification without sample preparation”

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Advantages

Compared to the current solutions which entail culture and susceptibility testing that can take days and a high throughput sequencing that requires expensive equipment and numerous technical steps, the benefits of LLNL’s MDx platform technology include:

  • Simple processing - sample is diluted and loaded – extensive sample prep not required
  • Rapid – results within 30 minutes
  • Low cost equipment – simple heating unit only, likely less than 25%
  • Simultaneous testing – pathogen and antibiotic resistant marker
Potential Applications

In addition to detecting and identifying biothreat agents and antibiotic-resistant, uropathogenic E. coli in urine, other applications envisioned are:

  • Food Safety
  • Plant pathogen detection
  • Animal pathogen detection
  • Human pathogen detection
  • Environmental sample testing
Development Status

LLNL is seeking a collaborative partnership to demonstrate and commercialize the capability of its prototype MDx platform to detect antibiotic-resistant, uropathogenic E. coli in urine. A focused study will demonstrate the utility of the system for pathogen detection in clinical settings.

LLNL has been testing assays with a collection of antibiotic-resistant E. coli strains with demonstrable success. The cartridge design is in final prototyping and testing before injection molding. LLNL has built and tested both a building-powered and battery-powered heating unit for these assays. LLNL holds issued patents on isothermal amplification techniques and applications.

LLNL’s patent portfolio includes:

  • US Patent App 14/257313 Multi-Chamber Nucleic Acid Amplification and Detection Device
  • US Patent App 13/767726 Isothermal Nucleic Acid Detection without Sample Preparation
  • US 8,808,643 Fluidics Platform and Method for Sample Preparation and Analysis
  • US Patent App 13/228,384 A Fluidics Cartridge and Reaction Platform
  • US Patent App 13/228,370 A Fluidics Platform and Method for Sample Preparation
  • US Patent App 13/115,877 Apparatus for Point-of-Care Detection of Nucleic Acid in a Sample
Reference Number
30816