PetChek HTWM PF

Heartworm Antigen Test Kit

For veterinary use only.

PetChek* HTWM PF

The PetChek* Heartworm Antigen Test is an enzyme immunoassay for the detection of Dirofilaria immitis (D. immitis) antigen in canine or feline serum or plasma.

A microtitration format has been devised in which antibodies to D. immitis antigen are coated on test wells. Upon incubation of the sample in the coated test well, antigen in the sample forms complexes with the coated antibodies. Following removal of the sample, an enzyme conjugated antibody is added and binds to antigen captured in the well. In the final step of the assay, unbound conjugate is washed away and enzyme substrate/chromogen is added. Subsequent color development indicates the presence of heartworm antigen in the sample.

Precautions and Warnings

● Do not expose TMB Substrate Solution to direct sunlight or any oxidizing agents.

● All wastes should be properly decontaminated prior to disposal.

● Do not mix components from kits with different serial numbers.

● Optimal results will be obtained by strict adherence to this procedure. Careful pipetting and washing technique throughout this procedure are necessary to maintain precision and accuracy.

● Refer to country specific Material Safety Data Sheet for regional hazard identification.

Storage

Store all reagents at 2-8°C. Allow reagents to come to room temperature (18-25°C) before use. Precipitation may occur in Stop Solution. Bring to room temperature (18-25°C) and mix by swirling before use.

Kit Components

Materials Required but Not Provided

● Distilled or deionized water

● Wash bottle

Use Of Precision Pipette To Dispense 100 µl

Place disposable pipette tip on pipette. Depress button on pipette, place tip in sample and release button slowly to pull up sample. Depress button to dispense sample in well.

Preparation Of Wash Solution

Dilute Wash Concentrate ten-fold (1/10) with distilled/deionized water (e.g. 5 mL of Wash Concentrate plus 45 mL of water for each 12 wells). Salt crystals may form in the Wash Concentrate upon storage. If this occurs, allow concentrate to come to room temperature (18-25°C) and mix by swirling to redissolve crystals before preparing wash solution.

Sample Information

Serum or plasma may be used in this test. Serum or plasma (e.g., Heparin, EDTA) may be stored for up to 7 days at 2-8°C. Highly hemolyzed samples should not be used, however moderately hemolyzed or lipemic samples will not affect results. For longer storage, sample should be frozen (-20°C or colder).

Test Procedure

Allow reagents to come to room temperature (18-25°C) before use. Mix reagents by gentle swirling.

1. Count the number of samples to be tested plus two wells for the positive and negative controls (one each). If using the well holder, remove the required number of wells from the bag and place into the well holder (up to 2 strips or 24 wells). If using the plate frame, remove the required number of complete strips from the bag and place into the 96-well plate frame. Record the positions of samples and controls on a worksheet.

2. Add 2 drops (100 µL) of Positive Control to the first well and 2 drops (100 µL) of Negative Control to the second well.

3. Using the precision pipette provided in kit and a separate pipette tip for each sample, add 100 µL of serum or plasma to appropriate wells. Incubate sample in well for 5 minutes at room temperature (18-25°C).

4. Discard the fluid from the wells by inverting. Slap firmly onto absorbent paper to remove all fluid.

5. Add 2 drops (70 µL) of HRPO Conjugate solution to each well. Incubate at room temperature (18-25°C) for 5 minutes.

6. Discard the fluid from the wells by inverting. Slap firmly onto absorbent paper to remove all fluid. Wash wells with a forceful stream of diluted wash solution by completely filling each well and discarding the fluid. Slap wells onto absorbent paper after each wash. Repeat for a total of 5 wash cycles.

7. Add 1 drop (50 µL) of TMB Substrate Solution to each well. Incubate at room temperature (18-25°C) for 5 minutes.

8. Add 1 drop (50 µL) of Stop Solution to each well. Mix by tapping gently. Read result visually. Color will be stable for 15 minutes.

Results

A sample is considered positive if it has more color than the negative control. For the assay to be valid, the positive control must develop a distinct blue color. Negative control must be clear or very lightly colored.

All positive samples that have not been run in duplicate should be retested. If duplicate samples or repeat testing yield inconsistent results, repeat test.

Warning

H317 / H402 / H412 / P261 / P280 / P333+P313

Conjugate / Negative Control / Positive Control - May cause an allergic skin reaction. Harmful to aquatic life. Harmful to aquatic life with long lasting effects. Avoid breathing mist/vapors. Wear protective gloves. If skin irritation or rash occurs: Get medical advice/attention.

H319 / P280 / P305+P351+P338 / P337+P313

Stop solution - Causes serious eye irritation. Wear eye protection/face protection. If in eyes: Rinse cautiously with water for several minutes. Remove contact lenses, if present and easy to do. Continue rinsing. If eye irritation persists: Get medical advice/attention.

Symbol descriptions

IDEXX Technical Support

USA/Canada: 1 800 248 2483 • idexx.com

Australia: 1300 44 33 99 • idexx.com.au

Europe: idexx.eu

VLN/PCN: 313/5018.02

*PetChek is a trademark or registered trademark of IDEXX Laboratories, Inc. or its affiliates in the United States and/or other countries.

Patent information: idexx.com/patents.

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