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White Paper on the Application of Molecular, Spectroscopic, and Other Novel Methods to Monitor Pathogens for Potable Reuse

Project: 14-17
RFP Open Date: July 9, 2015
RFP Close Date: August 13, 2015

Program: Principal
Funding Available: $50,000
In-Kind Requirement: At least 25% ($16,666) of the total project costs

Project Manager: Stefani McGregor


Rapid molecular measurement methods (such as methods incorporating quantitative polymerase chain reaction or qPCR) could be used for potable reuse pathogen and microbial indicator monitoring with same-day results. There is a large body of research on applications of molecular methods for water quality assessment but this has not been studied for potable reuse. Molecular methods could provide advantages relative to current culture-based methods, but this potential needs to be critically assessed.  Several logistical and technological challenges remain that make adoption of molecular methods difficult in potable reuse monitoring.

This white paper will focus on molecular methods, but will briefly address the challenges and opportunities in other novel approaches such as (but not limited to) spectroscopic, light scattering, flow cytometry, electrochemical (including surface plasmon resonance), mechanical (including cantilever and quartz crystal microbalance methods), and immunological detection methods.

Goals and Objectives

The project will develop a white paper assessing the application and need for using molecular and spectroscopy based methods as well as other novel methods for identification and quantification of pathogens in potable reuse facilities.  The white paper will assess the potential applications as well as limitations and challenges to implementation in a purified water matrix.

Research Approach

This project involves the development of a technical white paper documenting the current state-of-the-science on molecular characterization methods (such as qPCR, RT-qPCR, next-generation DNA sequencing or pyrosequencing, nucleic acid sequence based amplification (NASBA), and droplet digital PCR and other novel methods. These methods will be assessed for the potential to implement them to validate treatment performance and assess water quality with regards to microbial indicators and pathogens in potable reuse applications.   Key microbial groups of interest are enteric virus, bacteria, parasitic protozoa, and water-based (e.g., “opportunistic”) pathogens as well as conventional fecal indicators. Discussion should be provided how the applicability of the method varies depending on the target.


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