WITNESS EHRLICHIA

WITNESS EHRLICHIA

Canine monocytic ehrlichiosis is becoming one of the most important tick-born diseases. Caused by Gram negative bacteria Ehrlichia canis, the infection course is divided into three phases: acute, subclinical and then chronic in immunodepressed dogs. A true preva- lence of canine ehrlichiosis may be understated due to the tendency to either spontaneous recovery after the acute phase or death from superinfections in the chronic disease stage. In all cases, bacteriaemia can last for years and new clinical manifestations may arise.

 

TEST INDICATION

If the bacteria duplicate and persist in the host’s monocytes, direct identification on blood smears is hampered by the low number of these cells in blood. In general, the disease is diagnosed by the presence of specific antibodies; however some crossreac- tions with other pathogenic bacteria (Ehrlichia ewingii, E. risticii) have been reported.
The use of an antibody test in combination with others tests (blood cells count and blood smear examination for others pathogenic bac- teria) and the evaluation of clinical signs and recovery with an appropriate antibiotherapy, permits the correct diagnosis of canine monocytic ehrlichiosis .

The WITNESS® EHRLICHIA Antibody test for the detection of anti-Ehrlichia canis antibody is recommended for use when clinical history and/or clinical signs could suggest a canine ehrlichiosis.

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TEST PRINCIPLE

The WITNESS® EHRLICHIA Antibody test is a simple test, based on Rapid Immuno Migration (RIMTM) technology, using an antigen from E. canis to quickly identify antibodies in Ehrlichia- infected dogs. Sensitized colloidal gold particles bound to anti- E. canis antibodies present within the sample (whole blood, serum or plasma) are allowed to migrate along a strip. The complex is then captured on a sen- sitized reaction line where its accumulation causes the formation of a clearly visible pink band. A control band, located at the end of the reading window, ensures that the test was performed correctly.

SPECIMEN INFORMATION

• The test can be performed on unclotted whole blood anticoagulated with EDTA, sodium citrate or heparin, serum or plasma.
• Samples should always be collected with a sterile needle and syringe.
• Haemolysis does not significantly interfere with the test, but strongly haemolyzed samples may partly obscure a weak

positive line (due to haemoglobin background).

 

TEST PROCEDURE

Important: Allow samples and buffer drops to fall onto membrane at window (1). Avoid touching the membrane while applying the sample or buffer drops. Do not touch the membrane with pipette or buffer bottle tips.

1. Sample application

• Tear open a pouch provided and place the test device on a flat horizon- tal surface for the duration of the test.

• Holding the provided pipette vertically, transfer one drop of sample to the sample well , window (1).

2. Buffer dispensing

• Check that the sample has truly penetrated the membrane.
• Remove the cap from the buffer bottle, hold it vertically and add three drops of buffer to the sample well, window (1).
• Leave the test device flat during migration of sample/reagent complex through the reading window.

3. Reading test

• After 10 minutes, observe the presence or absence of pink / purple bands in reading windows (2) and (3).

• Sample results are read in window (2). The control band is read in win- dow (3).

Notes :

  • The test is complete and may be read before 10 minutes if two

    pink/purple bands are clearly visible in both windows (2) and (3).

  • The presence of a pink/purple band only in window (3) before the 10 minutes does not mean that the test is complete. A pink/purple band in window (2) may develop slower than the control band in window (3).

 

RESULTS
1.  Validation
Test is validated if a pink/purple band is present in the reading window (3). 2. Interpretation
• No band in reading window (2), with one band in window (3): sample is negative for E. canis antibodies
• One band in reading window (2), with one band in window (3): sample is positive for E. canis antibodies

Notes:

• No band in control window (3): invalid test.
•A test result should always be interpreted in the context of all available clinical information and history for the dog being tested.

  • It is recommended to re-test sample on serum or plasma in case of results that do not correlate with the clinical symptoms.
  • In case of doubtful results, it is advised to renew testing 1 or 2 weeks later.

 

Storage

It is preferable to test samples immediately after collection and not longer than 4 hours after collection, if stored at room temperature.
If testing is further delayed, samples should be refrigerated (+2°C – 8°C) for up to 2 days.
For prolonged storage, samples (serum and plasma only) should be kept frozen (-20°C).

 

KIT CONTENTS

A. 5 pouches,each containing 1test device and desiccant.

B. 1 Buffer dropper bottle (2ml).

C. Instructions for use.

D. 5 pipettes.

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