TM
VecTOR
MALARIA SPOROZOITE
ANTIGEN
PANEL ASSAY
TM
VecTOR
Malaria Sporozoite Antigen
Panel Assay
Prepared by
Entomological Sciences Division
USACHPPM-North
Fort George G. Meade, MD
The use or mention of a private entity in this document is not to be
construed or interpreted as an official federal endorsement of same.
Learning Objectives
Given a VecTOR test kit know:
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components
use
role in surveillance program
limitations
Malaria Adult Mosquito Surveillance Plan
Alternatives
• Do nothing
• Focus surveillance on
suspected risk areas
• Conduct tests in order
to predict malaria risk
Malaria Surveillance Considerations
• Vector
– Does it bite humans
– Can it transmit malaria to humans
– Is there a high vector abundance
• Environmental Factors
– Usually low to moderate illness level in endemic areas
– Epidemic outbreaks are likely when conditions are suitable
– Outbreaks can occur when climate is favorable for short periods
• Host
– Is there immediate onset of disease (10-20 days) or latent disease
manifestation (6-9 months - e.g. Korea)
– Is there low level immunity in host population
Adult Mosquito Surveillance
(Mosquito Magnet)
Adult Mosquito Surveillance
(Pickle Jar”, ABC, CDC and Shannon Traps)
Malaria Identification & Distribution
• Identify potential malaria high-risk areas
• Collect and identify adult mosquitoes
• Perform screening with VecTOR kit
– Follow up with PCR confirmation
• Define high risk areas
• Initiate targeted control operations
• Integrate into a GIS platform to facilitate an
effective management program
Malaria Transmission
Malaria VecTOR Kit
• Rapid immunochromatographic assay
for the qualitative detection of:
– Plasmodium falciparium
– Plasmodium vivax 210
– Plasmodium vivax 247
in infected mosquitoes
Malaria VecTOR Kit
• Results can enable PM organizations to:
–Continuously monitor mosquito vectors
–Focus vector control
–Deliver cost-effective disease monitoring
Malaria VecTOR Kit
• Intended to be a component of the Entomological
Collection Kit, Field (ECK, Field)
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Separate Bde PM Section (1)
Division PM Section (3)
ASMB PM Section (3)
Medical Detachment Preventive Medicine (3)
SF Group (1)
Civil Affairs Fld/UW BN
• Public Health Team (1)
• Civil Action Team (1)
Malaria VecTOR Kit
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Item: Test Kit, Malaria Detection (MAL-K020)
NSN: 6650-01-551-5327
Cage Code: 3M5F7; AAC: Local Purchase
Unit of Issue : Kit of 20 tests
Unit of Price: $260.00
– $13.00 per test (<10 female An spp. Mosquitoes/test)
• Shelf Life: Stable to expiration date when stored
at room temp (10-30 deg C or 50-86 deg F)
• Sensitivity: 92%; Specificity: 98.1%
• Estimated limit of detection: 200-400 sporozoites
DOD PEST MANAGEMENT MATERIEL LIST (OTHER THAN PESTICIDES)
June 1, 2007
Table of Contents
Section 1 - Hand Sprayer, Pesticide, Manually Operated, Compressed Air
Section 2 - Duster, Pesticide, Manually Carried
Section 3 - Sprayer and/or Duster, Pesticide, Manually Carried, Backpack
Section 4 - Sprayer, Hydraulic or Electric, Mist or Solid Stream, Pesticide, Pushcart
Mounted
Section 5 - Sprayer, Hydraulic or Electric, Mist or Solid Stream, Skid/Frame Mounted
Section 6 - Sprayer, Hydraulic or Electric, Mist or Solid Stream, Pesticide, Trailer Mount
Section 7 - Pesticide Sprayer, Fog/Aerosol Generator (ULV/ULD), Manually Carried
Section 8 - Pesticide Sprayer, Fog/Aerosol Generator (ULV/ULD), Skid Mounted
Section 9 - Pesticide Sprayer, Aerial Application Unit, Liquid or Dry Pesticide
Section 10 – Insect Traps, includes light traps and insect bait boxes
Section 11- Animal Traps
Section 12 - Rodent Bait Stations
Section 13 - Granular Spreader
Section 14 - Pheromone Trap Stored Products
Section 15 - Bird Pest Management Devices (Including bird traps)
Section 16 - Sets and Kits
Section 17 - Miscellaneous
Section 18 - Personal Protection Materiel
Section 19 - Devices for Electrocution of Flying Insects
Administrative Procedures (including emergency requisition of pesticides)
Malaria VecTOR Kit
– For Deployment Support contact the Deployment
Environmental Surveillance Program (DESP):
Commander, USACHPPM
5158 Blackhawk Road
ATTN: MCHB-TS-RDE
Aberdeen Proving Ground, MD 21010-5403
Phone: DSN 584-6096; FAX -2407
COM (410) 436-6096; FAX -2407
email: chppm-desprequests@amedd.army.mil
– Manufacturer Direct Orders contact:
VecTOR Test Systems, Inc.
2033 Yellowthroat Place
Thousand Oaks, CA 91320
Phone: (805) 390-0647, FAX (805) 376-8480
email: www.vectortest.com
Malaria VecTOR Kit
• Specimen Collection and Preparation
– Use appropriate collection methods to capture
female Anopheline species known to be malaria
vectors
– Collected Anopheles spp. should be immediately
identified and tested or,
– If not immediately tested, stored at -20 deg C until
they can be processed
Malaria VecTOR Kit
• Materials Provided
– Assay dipsticks (20) in canister with desiccant cap
– 1 Bottle of grinding
Solution
– 20 Conical Tubes
Malaria VecTOR Kit
• Materials Provided, Cont’d
– 2 Grinders
– 1 dropper cap
– 2 ten-hole tube
stands
Malaria VecTOR Kit
• Materials Required but Not Provided:
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Mosquito keys
Mosquito Collection/Pooling Log Sheet
Permanent Marker
Timing Device
Wash bottle
Motorized grinder (optional)
Malaria VecTOR Kit
• Materials Required but Not Provided:
– Forceps
– Latex gloves
– Coolers/gel packs
– White enamel or plastic sorting tray
– Dissecting Scope w/light source
Malaria VecTOR Kit
Grinding Solution:
– Before use:
• Mix gently, inverting the bottle five times
– When dispensing drops:
• Hold the bottle in an upside-down, vertical position
• Squeeze the white portion of the dropper cap just
above the the tip.
– Holding the bottle at an angle will cause the size of the
drops to vary, causing inconsistent solution dispensing
Malaria VecTOR Kit
• Procedure:
– Place 1-10 female Anopheline mosquitoes into
conical grinding tube
– Label tube with pool number
– Place the tube in the stand
– Dispense 13 drops of grinding solution into the
tube
Malaria VecTOR Kit
• Procedure Continued:
– Place pestle into the
grinding tube
– Rotate it vigorously to grind up the mosquitoes
– Wash the pestle
between samples (tubes)
Malaria VecTOR Kit
• Procedure Continued:
– Remove test strip (dipstick) from container
• Replace desiccant cap on canister to protect remaining test strips
– Label pool number
on the back side of
test strip
– Place test strip in
tube labeled with
corresponding pool
number
Malaria VecTOR Kit
• Procedure Continued:
Wait 15 minutes for test to be complete – read results
(Results should be read within 30 minutes. After 30 minutes dipsticks turn grayand results can be obscured.)
•DO NOT read dipsticks after they dry.
Malaria VecTOR Kit
• Procedure:
– Determine test results
• Remove test strip
• Compare test strip to pictorial sample in the
VecTOR Kit instructions
Presence of only a control
line indicates a negative
result
Test results are not valid if the
control line does not develop
Malaria VecTOR Kit
• Procedure:
• Presence one or more lines (other than control) indicates
the presence one or more malaria strains:
Plasmodium falciparum, P. vivax 210, or P. vivax 247.
Malaria VecTOR Kit
• Dipsticks can
be saved,
taped to a
sheet of paper
with pertinent
information added,
as a record of assay
results
Malaria VecTOR Kit
• Limitations:
– Can not test more than 10 mosquitoes per test
– Mosquito identification
• ID to genus vs. ID to species
– Screening Tool
• Can not be confirmed by PCR in a Laboratory from
strip (additional mosquito material is needed)
Malaria VecTOR Kit
• Used pipette tips, dipsticks, and sample
tubes can all be disposed of in regular
trash (all non-hazardous)
Malaria VecTOR Kit
• PM Dets, ASMB PM Sec, Div PM Sec, Sep Bde PM
Sec, SF Group PM, Civil Affairs Teams
– Another tool in your tool box
• Can be used to test field collections of Anopheline mosquitoes
– Hone your mosquito ID capability
– Practice Malaria vector surveillance
– Expect low infection rates, unless a large outbreak occurs
Malaria VecTOR Kit
Questions?
Malaria VecTOR Kit
– For Deployment Support contact the Deployment
Environmental Surveillance Program (DESP):
Commander, USACHPPM
5158 Blackhawk Road
ATTN: MCHB-TS-RDE
Aberdeen Proving Ground, MD 21010-5403
Phone: DSN 584-6096; FAX -2407
COM (410) 436-6096; FAX -2407
email: chppm-desprequests@amedd.army.mil
– Manufacturer Direct Orders contact:
VecTOR Test Systems, Inc.
2033 Yellowthroat Place
Thousand Oaks, CA 91320
Phone: (805) 390-0647, FAX (805) 376-8480
email: www.vectortest.com
Malaria VecTOR Kit
•
•
•
•
•
Item: Test Kit, Malaria Detection (MAL-K020)
NSN: 6650-01-551-5327
Cage Code: 3M5F7; AAC: Local Purchase
Unit of Issue : Kit of 20 tests
Unit of Price: $260.00
– $13.00 per test (<10 female An spp. Mosquitoes/test)
• Shelf Life: Stable to expiration date when stored
at room temp (10-30 deg C or 50-86 deg F)
• Sensitivity: 92%; Specificity: 98.1%
• Estimated limit of detection: 200-400 sporozoites
EXERCISE:
Using the Malaria Sporozoite
Antigen Panel Assay Kit
Objectives
Given field collected mosquitoes, a list of common
mosquito genera, a mosquito key, a Malaria
VecTOR Kit, DA Form 8011-R, Mosquito Pool
Log Sheet, a dissecting microscope w/light source:
• Sort Anopheline mosquitoes from all other genera
• Pool the Anopheline mosquitoes
• Test the pools using the VecTOR kit (malaria
sporozoite antigen panel assay)
Malaria VecTOR PE
CONDITIONS
You are supporting general mosquito (vectorborne disease surveillance) at an OCONUS base
camp. Given a country specific mosquito key,
dissecting microscope w/light source, field
collected mosquito specimens, DA Form 8011-R,
Malaria lab pooling sheet, Malaria VecTest
Antigen Panel Kit, permanent marker, and
appropriate label information, prepare specimens
for and test anopheline mosquitoes for malaria.
Malaria VecTOR PE
STANDARDS
1.
2.
3.
4.
Identify required materials
Properly record collection data on DA Form 80-11-R
Sort Anopheline mosquitoes from all other genera
Pool Anopheline mosquitoes by trap, by collection date; record
pool data on Malaria Pool Log Sheet
5. Test Anopheline pools using the VecTOR Kit
Follow VecTOR Kit directions:
Label vials
Label dip sticks
6. Record test results and archive test strips permit and/or labeling
requirements
7. Pack and label pool specimens shipment for shipment to a CONUS
Lab
Properly record
collection data on
DA Form 8011-R
MOSQUITO POOL INFORMATION - MALARIA SURVEILLANCE
POC name:
Location:
POC phone:
Country:
POC e-mail:
(+) = positive, (-) = negative
Pool
Collect.
Trap
Collect.
No. of
Log No.
Date
Site
Method
Mosq.
Mosquito ID
Notes/
Test
Comments
Date
Results
Pf
Pv210
Mosquito Pooling
Sheet
Pool Log No.: Should match label on vial (e.g., FGGM 001, FGGM 002, etc. for Fort Meade)
Collection Date:
mm/dd/yyyy. For consistency, use the date the traps were PUT OUT
Trap Site:
Use your own number/code scheme. Make sure you know how to link it to a site on map
Collection Method:
Light Trap w/CO2 (LTC), Light Trap w/out CO2 (LTX), Gravid Trap (GT),
Resting Collection (RC), Other (O) and specify in notes.
No. of Mosquitoes:
Total number within the pool. If zero, enter "0" but DO NOT ASSIGN A POOL # (use a "-")
Mosquito ID: Anopheles (An); if you know the species put it down
Test Date:
Date the dipstick assay was performed
Results:
Use plus symbol (+) to indicate a positive result and minus symbol (-) to indicate a negative result.
“Pf” = Pl. falciparum, “Pv 210”= Pl. vivax 210, “Pv 247” = Pl. vivax 247
Further Tips:
Only pool FEMALE mosquitoes. DO NOT USE DUPLICATE POOL NUMBERS.
Pv247