Faculty and Student Teams Program

Project Descriptions

Pacific Northwest National Laboratory
Biodefense Group

Development and Evaluation of Innovative Approaches for the Quantitative Assessment of Pathogens and Cyanobacteria and Their Toxins in Drinking Water

Requesting applications from science or engineering faculty members at institutions serving students underrepresented in science, engineering, mathematics and technology.

Project Description

Objectives: The overall objective of this project is to develop and demonstrate an automated system approach for large volume concentration, purification, and quantitative detection of CCL viruses in drinking water. For systems testing we will use bacteriophages MS2 and PRD1 as surrogates. Experimental approach:  Our research approach will comprise 5 Specific Aims. Specific aim 1: Evaluate the automated concentration of viruses from large volumes of drinking water using hollow fiber ultrafiltration (HFF) in the context of a) engineering into an automated sampling system, and b) providing efficient and reproducible concentration of viruses within an automated system. Specific aim 2: Develop and validate fluidic secondary capture and purification of intact viruses from the retained volumes in HFF to allow analysis by PCR and cell culture. Specific aim 3: Optimize fast, quantitative reverse transcription real-time PCR (qRT real-time PCR) for the detection of the purified hNoV in drinking water. Specific aim 4: Optimize our 3-D cell culture infectivity assay for hNoV, and test this assay for identification of infectious hNoV in spiked and unspiked drinking water samples. Specific aim 5: Design and test process integration of HFF and secondary capture and purification, and evaluate the system performance. For the purposes of this project, the concentrated samples will be split such that half of the purified sample will be evaluated offline by qRT real-time PCR and the other half evaluated for infectivity in the 3-D cell culture. Expected outcomes: Development of an approach for automated waterborne viral monitoring is expected to significantly improve reproducibility compared to manual methods. Secondary concentration using novel carbohydrate and lectin affinity reagents may provide broad agent secondary capture and purification such that the volumes delivered and evaluated in PCR represent 100 – 1,000 L of water.  The qRT real-time PCR detection component will provide a rapid (hours) and quantitative assessment of hNoV in drinking water; and novel 3-D cell culture techniques will allow, for the first time, evaluation of human NoV infectivity in environmental samples that will improve risk assessment models for this CCL virus.

FY2009 work with focus on Specific Aims 1 and 2.

Applicants Responsibilities and Relationship to Project

Applicants will receive support under the Department of Energy Faculty Student Team Research Program (FaST) to work collaboratively with the project research team at the Pacific Northwest National Laboratory for up to 10 weeks during the year starting in May/June. Summer and academic year visits to Pacific Northwest will be scheduled by mutual agreement between the Systems Risk and Analysis Group and the successful applicant. Ideally faculty will work at Pacific Northwest on the project for 10 weeks during the summer in the first year. Faculty will be expected to identify students from their campus to participate in the Undergraduate Research Participation programs offered by the Department of Energy at Pacific Northwest National Laboratory. Ideally Faculty will provide mentorship and/or advising support to students during the summer research activities. It is expected that the Faculty member will become an integral part of the research team working on this project and will support the project through the academic year on her or his campus.

Qualifications of Ideal Candidate

Support and Financial Commitments

See Financial Information.

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