MEGACOR FASTest BRUCELLA Canis Instruction Manual

MEGACOR FASTest BRUCELLA Canis

In vitro diagnosticum
Test-kit for the qualitative detection of Brucella canis IgG antibodies in whole blood, plasma or serum of the dog

INFORMATION ON THE TEST-KIT

TEST-KIT COMPONENTS

1. test-kit FASTest® BRUCELLA canis contains:
   • 2 or 10 test cassettes coated with recombinant Brucella canis antigens
   • 1 dropper bottle A with 1.0 ml or 3.0 ml buffer diluent
   • 2 or 10 disposable plastic pipettes
   • 1 instruction for use

LIABILITY
The entire risk due to the performance of this product is assumed by the purchaser. The manufacturer shall not be liable for indirect, special or consequential damages of any kind resulting from the use of this product.

ACCURACY
Sensitivity 72.7 %
Specifi city 90.7 %
(Comparison Method: Rose Bengal Test)

INTRODUCTION

• Brucella canis is a gram-negative bacterium appearing worldwide and being a potential danger for dogs and humans (zoonosis).

• Although the prevalences are very low or B. canis is partially seen rather obliterated in countries with high breeding standards, increased attention must be paid to brucellosis, especially in dog breedings. By mating with dogs from abroad (with lower breeding standards), brucellosis can be imported easily and unnoticed.

• The pathogen mainly propagates via infectious abort material or vaginal fl uids, by mating or vertically from bitch to pup-pies.

• Infected animals show failure in gravidity or infertility as well as atypical symptoms (e. g. uveitis). In about 75 % of the cases, females abort after 45 to 55 days of gestation. Early embryonic death and absorption or abortion 10 to 20 days after mating is reported, too. These abortions may go unnoticed, and the female then is often presented with the preliminary report “failure to conceive”.

• In males, the main signs are epididymitis, testicular atrophy and a moist scrotal dermatitis, in addition to bad semen quality (esp. with chronic brucellosis) and quality.
  Besides to missing or misunderstood symptoms, antibody levels in chronic animals can drop under the limit of detection. Hence, breeding dogs should be routinely tested for antibodies with serological methods to prevent the danger of propagation via venereal transmission.

• Being fast, simple and reliable, FASTest® BRUCELLA canis enables the veterinarian to have a complete on-site predication of the brucellosis status of the single animal or the complete breeding. Therapeutic and prevention measures can be applied immediately, adapted to dog and breeder needs.

INFORMATION ON THE SPECIMEN MATERIAL

• Approximately 25 µl (1 drop of attached plastic pipette) 15–25 °C warm whole blood (WB, with anticoagulant), plasma (P) or se-rum (S) are needed. Native blood without anticoagulant should not be used due to potential micro agglutination (e. g. migration delay on the membrane, unspecifi c reaction)!
  Mix the sample material well before use!

• Non-cooled (15–25 °C), WB, P and S should be tested within 4 hours! At 2–8 °C, WB, P and S can be stored up to 4 days. Plasma and / or serum samples can be permanently stored at minimum −20 °C.

• Keep in mind that the sample material, as well as all used test-kit components, should have reached room temperature at the time of application.

• Endogeneous and exogeneous interfering substances of the sample (e. g. albumin, fi brinogen, lipids, CRP, heterophilic anti-bodies, especially type IgA, as well as viscosity, pH-value and excess EDTA) as well as native blood can cause interferences (matrix effects) that can infl uence the target measurement. These can lead to an impaired LF and / or unspecifi c reactions on T and C.

SPECIMEN COLLECTION AND PREPARATION

• No specimen preparation necessary.
• ATTENTION: Partially fi lled and / or insuffi cient mixed EDTA, Citrate or Heparin tubes could create invisible mi-croclots resulting in lateral fl ow delay and / or unspecifi c reactions (e. g. greyish shadow like lines).

TEST PROCEDURE

1. Remove the test cassette from its foil pouch shortly before use. Place it on a fl at surface.
2. Express 1 drop (ca. 25 µl) of whole blood, plasma or serum into the sample window S using the disposable plastic pipette (fi g.1).
3. Hold the buffer dropper bottle A vertically and express 1 drop of buffer diluent (ca. 40–50 µl) into the sample window S (fi g.2).
4. Add 1 additional drop of buffer diluent into the sample window S if there is no beginning LF visible within 1 min-ute after adding the buffer diluent.

READING OF THE TEST RESULT

Read the test result 20 minutes after the drop of buffer diluent have been added into the sample window S.

POSITIVE TEST RESULT (fi g.3)
A pink-purple TEST line (T) of any intensity (varying from very weak to strongly intensive) and a pink-purple CONTROL line (C) appear.

NEGATIVE TEST RESULT (fi g.4)
Only a pink-purple CONTROL line appears. This line indicates, irrespective of its intensity, that the test has been performed properly.

INVALID TEST RESULT
No CONTROL line visible. The test should be repeated using a new test cassette.

PRECAUTIONS FOR USERS

• The guidelines for working in medical laboratories must be observed. It is recommended to wear disposable gloves and other personal protective equipment (protec-tive clothing, possibly a face mask). Wash and disinfect hands after completing the test.
• Label sample material and associated test cassette to ensure a precise assignment.
• Use a new pipette and a new test cassette for each sample.
• The buffer diluent contains low concentrations of toxic sodium azide as a preservative, therefore avoid skin/eye contact and/or ingestion.
• The sample material must be seen as potentially infectious and disposed of accordingly, together with the used test-kit components.

TEST PRINCIPLE

• The FASTest® BRUCELLA canis is based on an immuno-chromatographic “sandwich principle”.
• The Brucella canis antibodies of the sample will bind to specifi c mobile monoclonal antibodies, which are bound to gold particles. Migrating (“lateral fl ow”, LF) along the nitro-cellulose membrane, these antigen-antibody complexes are bound by specifi c recombinant B. canis antigens producing a pink-purple TEST line (T).
• A correct test procedure will be indicated by a second, pink-purple CONTROL line (C).

INFORMATION FOR THE INTERPRETATION

• The interpretation of the test result should always be based on anamnestic and clinical data as well as the therapy and prophylaxis possibilities.
• Any non-described colour or contour variation of T and C (e. g. greyish, shadow-like lines) or after more than 20 minutes has to be considered as unspecific reaction and therefore as negative test result.
• Due to anticoagulated whole blood and/or red hemoglobin background of the test membrane, caused by hemolytic blood samples, the visibility of T, especially in case of weak positive samples, could be from bad to not visible.

Positive test result

• Dog had contact with Brucella canis!
• To rule out whether the antibody reaction is based on an acute or chronic brucellosis, two serum samples at inter-vals of 2 to 4 weeks should be taken for testing with IFAT and / or Agar Gel Immunodiffusion (AGID) etc. A defi nite titre increase in the IFAT or AGID is indicative for an on-going Brucellosis.

Negative test result

• Dog had no contact with Brucella canis.
• Early brucellosis infection stage (< 2–4 weeks post infec-tion!). Dog has not yet produced antibodies in a detect-able concentration.
• In chronic infected animals, antibody titres cannot always be detected by single testing. Therefore, breeding dogs should be serologically tested routinely (multiple testing) to minimize the danger of venereal propagation of the pathogen.

Supplied Exclusively To The UK Veterinary Market By
Vetlab Supplies Ltd
Visit Our Website www.vetlabsupplies.co.uk
Telephone: 01798 874567
email us: [email protected]

References

• MEGACOR Diagnostik GmbH – Veterinary in vitro diagnostics
• Wholesale Vet Supplies & Products for Practices & Labs | Vet Supplies Online UK

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