VecTORVecTOR

MALARIA SPOROZOITE ANTIGEN

PANEL ASSAY

MALARIA SPOROZOITE ANTIGEN

PANEL ASSAY

TM

VecTOR

Malaria Sporozoite AntigenPanel Assay

Entomological Sciences DivisionUSACHPPM-North

Fort George G. Meade, MD

TM

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.

Learning Objectives

Given a VecTOR test kit know:

– 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

• Results can enable PM organizations to:

–Continuously monitor mosquito vectors

–Focus vector control

–Deliver cost-effective disease monitoring

Malaria VecTOR Kit

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)

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

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:

– 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

• 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.

Malaria VecTOR Kit

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

• 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

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)

• Used pipette tips, dipsticks, and sample

tubes can all be disposed of in regular

trash (all non-hazardous)

Malaria VecTOR Kit

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

CONDITIONSYou 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.

Malaria VecTOR PE

STANDARDS1. Identify required materials2. Properly record collection data on DA Form 80-11-R3. Sort Anopheline mosquitoes from all other genera4. Pool Anopheline mosquitoes by trap, by collection date; record

pool data on Malaria Pool Log Sheet5. Test Anopheline pools using the VecTOR Kit

Follow VecTOR Kit directions:Label vialsLabel 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 Pooling Sheet

MOSQUITO POOL INFORMATION - MALARIA SURVEILLANCE

POC name:

Location: POC phone:

Country: POC e-mail:

(+) = positive, (-) = negative

Pool Collect. Trap Collect. No. of Notes/ Test Results

Log No. Date Site Method Mosq. Mosquito ID Comments Date Pf Pv210 Pv247

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.