1. MALARIA SPOROZOITE ANTIGEN PANEL ASSAY
VecTOR TM
MALARIA SPOROZOITE ANTIGEN
PANEL ASSAY

2. VecTOR Malaria Sporozoite Antigen Panel AssayTM
Malaria Sporozoite Antigen
Panel Assay
Entomological Sciences Division
USACHPPM-North
Fort George G. Meade, MD
Prepared by
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.

3. Learning Objectives Given a VecTOR test kit know: components use
role in surveillance program
limitations

4. Malaria Adult Mosquito Surveillance Plan Alternatives
Do nothing
Focus surveillance on suspected risk areas
Conduct tests in order to predict malaria risk

5. 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

6. Adult Mosquito Surveillance (Mosquito Magnet)

7. Adult Mosquito Surveillance (Pickle Jar”, ABC, CDC and Shannon Traps)

8. 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
Suspected malaria high-risk areas can be identified by monitoring human populations (especially civilian) infected with malaria. Adult Anopheles mosquitoes from surveillance collections for identifying vector/disease control strategies can be assayed by ELISA to determine the mosquito infection rate for vivax malaria parasites. These data will establish baseline data and provide estimates of risk of malaria transmission.
When integrated into the GIS system, malaria infection rates can be correlated with larval habitats, adult mosquito populations and human populations for developing predictions and trends in malaria transmission.

9. Malaria Transmission

10. Malaria VecTOR Kit Rapid immunochromatographic assay
for the qualitative detection of:
Plasmodium falciparium
Plasmodium vivax 210
Plasmodium vivax 247
in infected mosquitoes

11. Malaria VecTOR Kit Results can enable PM organizations to:
Continuously monitor mosquito vectors
Focus vector control
Deliver cost-effective disease monitoring

12. Malaria VecTOR Kit
Intended to be a component of the Entomological Collection Kit, Field (ECK, Field)
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)

13. Malaria VecTOR Kit Item: Test Kit, Malaria Detection (MAL-K020)
NSN:
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 deg F)
Sensitivity: 92%; Specificity: 98.1%
Estimated limit of detection: sporozoites

14. 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)

16. 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
Phone: DSN ; FAX -2407
COM (410) ; FAX -2407
Manufacturer Direct Orders contact:
VecTOR Test Systems, Inc.
2033 Yellowthroat Place
Thousand Oaks, CA 91320
Phone: (805) , FAX (805)

17. 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

18. Malaria VecTOR Kit Materials Provided
Assay dipsticks (20) in canister with desiccant cap
1 Bottle of grinding
Solution
20 Conical Tubes

19. Malaria VecTOR Kit Materials Provided, Cont’d 2 Grinders 1 dropper cap
2 ten-hole tube
stands

20. Malaria VecTOR Kit Materials Required but Not Provided: Mosquito keys
Mosquito Collection/Pooling Log Sheet
Permanent Marker
Timing Device
Wash bottle
Motorized grinder (optional)

21. 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

22. 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

23. 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

24. 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)

25. 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

26. 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 gray- and results can be obscured.)
DO NOT read dipsticks after they dry.

27. 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

28. 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.

29. Malaria VecTOR Kit Dipsticks can be saved, taped to a sheet of paper
with pertinent
information added,
as a record of assay
results

31. 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)

32. Malaria VecTOR Kit Used pipette tips, dipsticks, and sample
tubes can all be disposed of in regular
trash (all non-hazardous)

33. 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

34. Malaria VecTOR Kit
Questions?

35. 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
Phone: DSN ; FAX -2407
COM (410) ; FAX -2407
Manufacturer Direct Orders contact:
VecTOR Test Systems, Inc.
2033 Yellowthroat Place
Thousand Oaks, CA 91320
Phone: (805) , FAX (805)

36. Malaria VecTOR Kit Item: Test Kit, Malaria Detection (MAL-K020)
NSN:
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 deg F)
Sensitivity: 92%; Specificity: 98.1%
Estimated limit of detection: sporozoites

37. Using the Malaria Sporozoite Antigen Panel Assay Kit
EXERCISE:
Using the Malaria Sporozoite
Antigen Panel Assay Kit

38. 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)

39. Malaria VecTOR PE CONDITIONS
You are supporting general mosquito (vector-borne 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.

40. Malaria VecTOR PE STANDARDS 1. Identify required materials
2. Properly record collection data on DA Form R
3. Sort Anopheline mosquitoes from all other genera
4. 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

41. Properly record collection data on
DA Form 8011-R

42. Mosquito Pooling
Sheet