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Increasing the Capacity of US Public Health &
WLA Labs to Detect Biothreat Agents in
Drinking Water
Vincent Hill, Environmental Engineer
NCEZID, DFWED, Waterborne Disease Prevention Branch
Water Laboratory Alliance (WLA) Security Summit,
March 23, 2012
National Center for Emerging and Zoonotic Infectious Diseases
Division of Foodborne, Waterborne and Environmental Diseases
Background
US drinking water systems vulnerable to intentional
contamination
Also source water, treatment, and distribution system deficiencies;
premise plumbing issues
Priority for public health: Emergency preparedness
Ideally, use “universal” method to recover unknown biothreat
agent or multiple agents of concern
“Ultrafiltration” (UF) fits the bill
Pore sizes rated on molecular scale (~1-10 nm)
Can recover viruses, bacteria, parasites—even large toxins
Commercially available, relatively inexpensive “dialyzers”
Hollow Fiber UF Schematic
Water Sample
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Microbes
Suspended solids
Colloidal matter
Large dissolved molecules
UF Membrane
Filtered Water
(aka “filtrate” or “permeate”)
EPA & CDC Collaborating for Water-Related
Emergency Preparedness
Began collaborating ~2003 on UF methods
EPA’s focus: field-deployable instrument
Automated UF device developed
CDC’s focus: lab-based method for Laboratory
Response Network (LRN)
Lab-based UF method and associated secondary processing
protocol (“LRN Water Processing Protocol”) posted for LRN lab use
in 2007
EPA-CDC Study: No overall significant difference in
microbial recovery efficiency between the methods
(EPA 600/R-11/103, October 2011)
LRN Water Processing Protocol
10-100 L Drinking Water Sample
Ultrafiltration
Centrifugal Ultrafilter
(viruses, toxins)
Nucleic Acid
Extraction
Virus Testing
(e.g., qPCR)
Toxin Testing
Membrane filtration
(bacteria)
Culture
Direct
Nucleic Acid
Extraction
Boil PrepqPCR
qPCR
Tangential-flow Ultrafiltration
Centrifugal Ultrafiltration
Microfiltration
Proficiency Testing Project
Primary Goal: Increase number of LRN and WLA labs
identified by LRN as proficient to perform the protocol
Secondary benefit: Increasing capacity to perform large-volume
water sample processing for waterborne pathogens
Enrolled 15 labs
Workshop and hands-on training at CDC (Atlanta)
Method practice at home lab
Proficiency Testing Study: April-May 2012
Focus: biothreat agent recovery and detection
Also: Use of new Quality Control parameter (E. faecalis recovery %)
Acknowledgements
EPA Collaborators
Latisha Mapp
Prisca Takundwa
Erin Silvestri
LRN Colleagues
Stephen Morse
Beth Schweitzer
Laura Jevitt
CDC Project Staff
Chandra Schneeberger
Jackie Knee
Bonnie Mull
"The findings and conclusions in
this presentation have not been
formally disseminated by CDC
and should not be construed to
represent any agency
determination or policy"
National Center for Emerging and Zoonotic Infectious Diseases
Division of Foodborne, Waterborne and Environmental Diseases