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PCR-Based Pathogen Detection in Water

Technology #20160100

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Researchers
Satoshi Ishii, PhD
Assistant Professor, Soil, Water, and Climate; BioTechnology Institute
External Link (www.swac.umn.edu)
Managed By
Larry Micek
Technology Licensing Officer 612-624-9568
Patent Protection

US Patent Pending

Pathogenic Microbe Measurement

A genetically engineered microbe can be used as a process control strain to accurately determine concentrations of common pathogenic microbes in a water sample. This method begins by adding known amount of a process control strain of genetically-modified, non-pathogenic Pseudogulbenkiania bacteria to an environmental (e.g., water) or clinical sample. Primers and probe sequences are also added to amplify the DNA of both the genetically engineered microbe and the test microbe. An assay based on polymerase chain reaction (PCR) extracts amplifies the DNA of microbes present, thus determining the DNA recovery efficiency of the genetically engineered microbe and calculating the number of cells of the test microbe in the sample.

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Water Quality Assessment

Accurately detecting pathogenic microbes in water sources is critical in preventing adverse effects to human health and water ecosystems. Current methods that use fecal indicator bacteria such as Escherichia coli and enterococci to monitor levels of fecal contamination in waterways are prone to the environmental survival and growth of these bacteria, which limit their reliability in indicating contamination. Molecular tools such as PCR and quantitative PCR can be used to detect and quantify pathogenic bacteria, but these reactions may be inhibited by humic substances commonly present in soil and sediment, resulting in inaccurate results. Furthermore, some portions of DNA and/or microbial cells can be lost during testing, resulting in reduced DNA recovery efficiencies. This technology standardizes DNA extraction efficiencies for pathogen detection and yields accurate measurements of pathogenic microbe concentrations.

BENEFITS AND FEATURES:

  • Microbe-based biosensor to monitor water quality
  • Accurate microbial concentration determination
  • Process control strain of genetically-modified, non-pathogenic Pseudogulbenkiania bacteria
  • Assay based on polymerase chain reaction (PCR)

APPLICATIONS:

  • Water testing
  • Water quality monitoring
  • Could be applied to other environmental parameters such as food, soil, and fertilizer
  • Potential applications in biomedical settings to quantify contamination of biological samples

Phase of Development - Prototype Developed. Applied to water quality monitoring.