Development and Evaluation of an Innovative System for the Concentration and Quantitative Detection of CCL Pathogens in Drinking WaterEPA Grant Number: R833003
Title: Development and Evaluation of an Innovative System for the Concentration and Quantitative Detection of CCL Pathogens in Drinking Water
Investigators: Tzipori, Saul , Walt, David , Zuckermann, Udi
Institution: Tufts University
EPA Project Officer: Klieforth, Barbara I
Project Period: August 1, 2006 through August 1, 2009 (Extended to July 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: Water , Drinking Water
This proposal will develop, evaluate and validate a rapid method for the quantitative assessment of pathogens/indicator organisms from large volumes of source and drinking water. Microorganisms of concern will include protozoa (Cryptosporidium, Giardia, Microsporidium), bacteria (E. coli, Legionella, Helicobacter pylori, Aeromonas hydrophila, Micobacterium avium intracellular), algae (Cyanobacteria), and viruses (Adenoviruses, Caliciviruses, Coaxackieviruses, Echoviruses). We propose integrating the technologies of concentration, purification, detection and testing for viability/infectivity into a universal simplified economic and user friendly methodology.
Concentration of the microorganisms will be by continuous flow centrifugation (CFC) and the detection of a small number of microorganisms from large volumes of water will be done using multiplex miniaturized fiber bead microarrays. In a previous EPA STAR award we developed, optimized and validated a novel method, the CFC, for the concentration of three protozoa (Cryptosporidium spp. Giardia, Microsporidia) from large volumes of water (1000L). In this award we propose to expand the CFC approach to include selected Contaminant Candidate List (CCL) pathogens, including bacteria, algae and viruses. The proposed method will integrate the concentration of protozoa, bacteria, algae and viruses from water into a single concentration procedure using a continuous flow and compare it with conventional methods. We will focus on detection and quantitative identification of CCL pathogens in water using multiplex miniaturized fiber optic DNA bead microarrays coupled with a compact confocal-type imaging system. The proposed detection technique will be rapid, cost-effective, portable, robust and employ simple procedures.
The major tasks involved with this approach include: (1) integrate the concentration of protozoa, bacteria, algae and viruses from water into a single concentration procedure; (2) select sequences, develop assays, prepare bead microarrays, and test both synthetic and spiked samples; and (3) validate the combined method with complex water matrices.
At the completion of laboratory and field validation studies we anticipate that the proposed approach will enable the detection of small numbers of waterborne pathogens from large volumes of various water matrices in less than four hours.