Rapid and Quantitative Detection of Helicobacter Pylori and E. Coli O157 in Well Water Using a Nano-Wired Biosensor and QPCREPA Grant Number: R833005
Title: Rapid and Quantitative Detection of Helicobacter Pylori and E. Coli O157 in Well Water Using a Nano-Wired Biosensor and QPCR
Investigators: Alocilja, Evangelyn C. , Rose, Joan B. , Molloy, Stephanie L.
Current Investigators: Alocilja, Evangelyn C. , Rose, Joan B. , Dreelin, Erin
Institution: Michigan State University
EPA Project Officer: Hiscock, Michael
Project Period: November 1, 2006 through October 31, 2009 (Extended to October 31, 2011)
Project Amount: $600,000
RFA: Development and Evaluation of Innovative Approaches for the Quantitative Assessment of Pathogens in Drinking Water (2005) RFA Text | Recipients Lists
Research Category: Drinking Water , Water
The hypothesis of this proposal is that a disposable biosensor and QPCR can be combined seamlessly to develop a unique biosensor-QPCR as a tool for near real-time determination of contaminant occurrence in drinking water. The specific objectives are: (1) Develop a protocol for processing water samples for the biosensor and QPCR; (2) Assess the performance of the biosensor and QPCR for sensitivity, specificity, recovery, and false positives/negatives of detection and enumeration for E. coli O157:H7 and H. pylori in groundwater samples from the field; (3) Develop a method for detecting and enumerating E. coli O157:H7 and H. pylori by QPCR using bacteria isolated and screened by the biosensor system; and (4) Validate a method for testing viability of E. coli O157:H7.
The first step in our experimental approach is to evaluate the performance of the biosensor and QPCR separately. Once confirmed, a method to seamlessly integrate these two devices into a biosensor-to-QPCR field-lab technique, coupled with a viability test, will be developed.
This project is expected to advance the use of antibody-based methods and molecular techniques for application to drinking water supplies. The expected deliverables are: (a) proof of concept and assessment of biosensor and QPCR techniques for recovery, detection and quantitation of bacteria in water; and (b) the development of a unique attribute that is not in the literature at the moment: the capability for direct confirmation through QPCR of presumptive results from the biosensor, thereby minimizing false positive and false negative analysis.