The importance of fast, reliable detection of pathogens in drinking water in the United States and worldwide, is increasing rapidly due to rising populations and increasing environmental degradation, as well as the threat of terrorism. Although the technology readiness level of rapid detection methods for pathogens in water has increased dramatically in recent years, rapid, automated sample concentration and preparation have lagged significantly. The program proposed herein will combine a novel, rapid hydrosol concentrator with proven sample preparation methods and quantitative polymerase chain reaction (qPCR) to provide a detection system for pathogens in drinking water of unparalleled speed, reliability, and significantly lower detection limits. The InnovaPrep hydrosol concentrator was developed through a partnership of two Missouri companies, AlburtyLab, Inc., and Page Applied Research LLC, to quickly and efficiently concentrate waterborne bacteria, viruses, toxins, protozoa, or other particles of interest from a large liquid volume into a very small liquid volume. The InnovaPrep system uses a novel elution method to recover organisms that have been captured onto hollow fiber ultrafiltration membranes into very small volumes. AlburtyLab proposed development of a two-staged unit for rapidly concentrating volumes of 10 to 100 liters of water into volumes as small as 500 microliters in less than 1 hour. Two methods will be evaluated for removal of inhibitors and further concentration of the InnovaPrep output. The first method will use immunomagnetic beads, prior to lysis, to separate target organisms. The second method will use Boreal GenomicsÂ’ electrophoretic-based SCODA system, post lysis, to separate nucleic acids. A final analysis-ready sample of 50 microliters will be analyzed by qPCR. The anticipated results of the program are the development of an automated system that can provide rapid, automated, highly efficient concentration and preparation of nucleic acids from waterborne microbial pathogens into microliter-range volumes that can be directly analyzed by new, rapid microbiological methods. Potential commercial applications include use in: water security monitoring, quality control detection systems for water treatment plants, drinking water laboratories for automated processing of water samples, and automated microorganism detection systems for ultrapure water systems. Supplemental
Keywords: small business, SBIR, EPA, homeland security, drinking water pathogens, hydrosol concentrator, quantitative polymerase chain reaction, qPCR, drinking water, microbiological methods, water security monitoring, automated microorganism detection systems, microbial pathogens measurement, rapid detection methods
