PCR-Based Pathogen Detection in Water

Pathogenic Microbe MeasurementA 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 Pseudogulbenkianiabacteria 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.Water Quality AssessmentAccurately 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 qualityAccurate microbial concentration determinationProcess control strain of genetically-modified, non-pathogenic Pseudogulbenkiania bacteriaAssay based on polymerase chain reaction (PCR)APPLICATIONS:Water testingWater quality monitoringCould be applied to other environmental parameters such as food, soil, and fertilizerPotential applications in biomedical settings to quantify contamination of biological samplesPhase of Development - Prototype Developed. Applied to water quality monitoring.

USA