Support of Far-Forward Disease Surveillance
Operations with Deployable, Real-Time
Vector-Borne Disease Agent Analytic
Capability
February, 2010
COL JAMES SWABY
Chief Clinical Research Division
Wilford Hall Medical Center
Comm: (210) 292-7068)
DSN: 554-7068
email: james.swaby@Lackland.af.mil
Vector Surveillance Analytic System (VSAS)
Disease agent detection platform:
- Portable (one person)
Dengue,Thailand 2007
- Field-durable
- Field-sustainable
- Real-time PCR
- RAPID
JBAIDS
Leish, Kenya 2008
RAPID
Operating off Car Battery
Far forward
disease surveillance
Rugadized Advanced Pathogen
Identification Devise
RAPID
Origins
RAPID Used in CBAWS
- Chemical & Biological Detection System
- Detector Alarmed
- Detector Collected
Sample
- Individual Collected
Filter
- Sample Taken to Lab
- Real-time PCR
- RAPID
Origins
Dengue, Honduras 2002
First target: Dengue
Cutaneous Leishmaniasis
Tallil AB, Iraq 2003
Redirected to
Leishmaniasis
Control
Program
Photos – LTC Weina
Operational Application
Leish. surveillance support:
- USA Prev. Medicine &
- USAF Pub Health
Leish,Tallil AB, Iraq 2003
Leish,Tallil AB, Iraq 2003
Surveillance
Control
Force protection support:
Iraq:
Afghanistan:
- Tallil AB
- Kabul
- Camp Victory
- Balad AB
- Kirkuk AB
Operations
Operates
as:
.
- Stand alone field surveillance activity
- Extension of deployable assets:
-- USA TAML
-- AF BAT
Tallil AB, Iraq 2003
USA TAML
Hanta Virus 2003
AF BAT
Field Testing
Proven in diverse operations & environmental conditions:
- Leishmania Southwest Asia, 2003, 2004
- Leishmania Kenya, 2009, 2010
- Dengue, JTF Bravo, Honduras 2002
- Dengue, AFRIMS, Thailand 2003, 2007, 2008
- Malaria, AFRIMS, Thailand 2008
- Genomics-based id. Haughton-Mars Project, NASA,
Devon Island, Arctic
Devon Island, Arctic
NASA Haughton-Mars Project
Lineage
JPEO-CBD requirement = ID. Tech for ops. diseases.
- JPEO threat agent list
- Includes Dengue (block 1, tier 2) & Leish.
- Adopted JBAID analytic system
- Incrementally establishing VSAS as component of
JBAID
RAPID
JBAIDS
Light Cycler
Components
- Assays (freeze dried): field-durable, real-time
-
Positive controls
Negative controls
Preformatted reagents
PCR water
RAPID or JBAIDS
ITI reagent kit color coded
simplifies preparation
Extract Purification Kit
- Commercial off the shelf
-- Thermally stable
-- Preformatted
-- No 4o C cold chain
-- RAPID mini centrifuge
-- Adopted for:
--- Field-worthiness
--- Applicable to RNA & DNA
-- AVL lysis buffer
--- Inactivates agents
--- RNA remains stable days to weeks
-- Kit purifies nucleic acid from diverse matrices
--- Mammalian fluids/tissues
--- Arthropods homogenate
Assays
-
Assay primers & probes designed de novo
Freeze-dried reagents made by ITI®
Standardized for PCR and RT-PCR
Assays only require hydration & sample for analysis
Thermal stability:
-- Eliminates:
--- -20o cold chain,
--- Cold storage
--- Resupply issues
-- Enhances mobility
RAPID
- Commercialized version of JBAIDS
-- Assays easily transitioned
-- Real-time fluorimeter
-- Closed capillary design
-- 32 sample capacity
-- Laptop runs PCR cycling
-- Automated data
management
-- Mini Centrifuge for:
--- Sample prep
--- Capillary loading
Laptop
RAPID
Centrifuge
Field Durable
- Fits in 2 hardened & waterproofed cases
-- RAPID
-- Several hundred assays
-- Processing supplies
-- Power converter
-- Power cables
One man transportable
- Transportation
-- Personal equipment
-- Vehicle
-- Helicopter
-- Fixed Wing (baggage)
Field Durable
- Small footprint
-- Hand-carried
-- Vehicle tailgate
- Power sources
-- 110 & 220V AC
--- Fixed structure
--- Generator
Dengue, Thailand 2007
Generator
RAPID
Malaria, Thailand 2008
110 – 220 V AC
VSAS Field Durable
Dengue, Thailand 2007
- Power sources
-- 12V DC
Converter
Dengue, Thailand 2008
Running vehicle
Battery
Field Durable
- Easy set up & operation
-- Configured in 10 min.
-- Processing & analysis 2 hrs.
-- Sample processing &
master mix prep.
--- Preformatted reagents
--- Sample prep. Kits
--- Closed capillary design
-- No hood
-- No spatial separation
Malaria, Thailand 2008
Easy set up
Field Durable
Malaria, Thailand 2008
- Safety
-- Knowledge of vectors
-- Skill
-- Personal protective equip.
Skill
Knowledge
Safety gear
Dengue, Honduras 2002
VSAS Provides
“Real-time” Risk Assessment
- Affects time-critical & focused measures:
-- Prevention
-- Control
-- Most effective approach
Dengue
Yellow Fever
Aedes aegypti
- Reduce risk
Breeding site
-- Focus on high risk areas
Dengue, Honduras 2002
--- Infected vectors
--- Breeding Populations
VSAS Provides
Deployable Real-time Diagnostics
- To effect disease:
-- Risk assessment
-- Prevention
-- Control
Leish,Tallil AB, Iraq 2003
Assessment
Leish,Tallil AB, Iraq 2003
Prevention
Control
Products To Date
- Concurrently submitting to JPEO - JBAIDS
for FDA approval
-- Dengue Universal already in testing
- AFPMB Approved Surveillance Assays
-- Leish.:
--- LEIS
--- LVL
-- Dengue
--- DU-JCM
- DOD ID Protection
-- LEIS (D&RI)
-- LVL (D&RI)
-- Dengue (pending)
Leish, Kenya 2008
- 6 Publications
Currently Status
Diarrheal Diseases
Assay
E. coli enterotoxigenic (ETEC)
Heat Labile Toxin (LT)
Heat Stable Toxin (ST1a)
Heat Stable Toxin (ST1b)
Shigella spp
Cryptosporidium spp
Norovirus
*Global Risk Index
Rank*
2
2
2
2
12
N/A
Activities Underway
Comments
JPO/JBAIDS Sub Prep Dried Assay Validated
JPO/JBAIDS Sub Prep Dried Assay Validated
JPO/JBAIDS Sub Prep Dried Assay Validated
Dried Assay Validation Wet Assay Optimized
Dried Assay Production Wet Assay Optimized
Wet Assay Optimization Funded FY10
Currently Status
Vectorborne Diseases
Assay
Dengue virus
Chikungunya virus
Aedes aegypti mosquitoe
Leishmania spp
Leishmania visceral genotype
Leishmania human pathogenic
Leptospira human pathogenic
Leptospira spp
Plasmodium spp, P. f and P. v
Flavivirus spp
Japanese enchephalitis virus
*Global Risk Index
(Burnette et al., 2008)
Rank*
3
6
N/A
30
20
various
7
7
1
various
24
Activities Underway
Deployed Surveillance
Wet Assay Optimization
Deployed Surveillance
Deployed Surveillance
Deployed Surveillance
Wet Assay Optimization
Dried Assay Validation
Dried Assay Production
Wet Assay Optimization
Wet Assay Optimization
Wet Assay Optimization
Comments
AFPMB Approved
Funded FY10
AFPMB Sub Planned
AFPMB Approved
AFPMB Approved
Funded FY10
Field Val Planned
Wet Assay Optimized
Funded FY10
Funded FY10
Funded FY10
Future
- 2010 Grants, >$700K, for:
-- Enhanced Plasmodium genus assay
-- Leishmaniasis panel
-- Japanese Encephalitis
-- Sticky Ovitrap:
--- Chikungunya
--- Dengue
-- Norovirus
Malaria, Thailand 2008
Joint Effort
- Air Force:
-- SGR
-- 59 CRD
- Army:
-- WRAIR
-- AFRIMS
-- USAMRU-K
- Navy: NMRC
- ITI via CRADA