MEGACOR EHRLICHIA FASTest ANAPLASMA Instructions

MEGACOR EHRLICHIA FASTest ANAPLASMA

Test-kit for the qualitative detection of antibodies against Anaplasma spp. (A. phagocytophilum,
A. platys) and against Ehrlichia canis in whole blood, plasma or serum of the dog

INSTRUCTIONS FOR USE

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

Manufacturer:
6912 Hörbranz – AUSTRIA www.megacor.com

 INFORMATION ON THE TEST-KIT

TEST-KIT COMPONENTS
test-kit FASTest® ANAPLASMA-EHRLICHIA contains:

• 2 or 10 twin test cassettes coated with recombinant Ana-plasma spp. or Ehrlichia canis antigens
• 1 dropper bottle A with 1.0 ml or 3.0 ml buffer diluent
• 2 or 10 disposable plastic pipettes
• 1 instructions 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.

2. INTRODUCTION

• Canine Granulocytic Anaplasmosis (GA, Anaplasma phagocy-tophilum [A. p.], Thrombocytic Anaplasmosis (TA, Anaplasma platys) and Canine Monocytic Ehrlichiosis (CME, Ehrlichia canis) belong to the most important vector-borne infectious diseases induced by canine parasites. Co-infections play a major role in both pathogens, therefore, depending on the animals’ place of origin and whereabouts, potential other pathogens (e. g. Borrelia spp., Babesia spp., Leishmania spp.) or laboratory values indica-tive of these pathogens (e. g. anaemia, leucocytosis / neutrophilia, hypoalbuminaemia / hypergammaglobulinaemia, kidney values etc.) should be clarifi ed.
• In principle, tick territories (endemic area) are potential breeding grounds. Seroprevalences for GA / TA and CME vary widely, de-pending on country (endemic or non-endemic) and study. With an incubation time of 2–20 days, A. phagocytophilum in-fections are often subclinical or self-limiting. Clinical symptoms are fever, apathy, stiff muscle, polyarthritis with joint pain / swell-ing, lameness, weight loss, thrombocytopenia, anaemia, pete-chial haemorrhages and increasing infl ammatory values (CRP, haptoglobin). Subarachnoid haemorrhage could lead to central nervous disorders. The cause of TA is A. platys. It occurs world-wide, mainly in the Southern Hemisphere. In most cases, the infection is asymptomatic with mild fever, uveitis, petechiae and ecchymoses. In the laboratory diagnostics, a thrombocytopenia is shown.
• CME is characterised by a very long incubation period (Ø 4–5 up to 12–13 years) and an unspecifi c clinic. It is therefore also referred to as a “silent killer”!
  Very few veterinarians are aware of the consequences of such a co-infection from an immunological, therapeutic and diagnostic point of view. Therefore, FASTest® ANAPLASMA-EHRLICHIA is suitable as a quick, qualitative antibody (ab) detection if an ana-plasmosis, ehrlichiosis or a co-infection is suspected. This ena-bles to start immediately further diagnostic investigations as well as therapeutic and prophylactic measures.

INFORMATION ON THE SPECIMEN MATERIAL

• 10 µl each (1 drop of attached plastic pipette) 15–25 °C warm whole blood (WB, with anticoagulant), plasma (P) or serum (S) are needed. Native blood without anticoagulant must not be used due to potential micro agglutination (e. g. migration delay on the membrane, unspecifi c reac-tion)!
  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. Serum and / or plasma 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 tempera-ture at the time of application.

• Endogeneous and exogeneous interfering substances of the sample (e. g. albumin, fi brinogen, lipids, CRP, hetero-philic antibodies, 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 twin test cassette from its foil pouch shortly before use. Place it on a fl at surface.
2. Draw sample up to the mark (=^ 10 µl sample volume) using the disposable plastic pipette. Place the whole sample volume (10 µl) into the sample window S of the Ana-plasma test cassette (hold pipette vertically, fi g.1). Re-peat the whole procedure for the Ehrlichia test cassette.
3. Hold the dropper bottle A vertically and place 2 drops (ca. 80–100 µl) of the buffer diluent into the sample window S of the Anaplasma and the Ehrlichia test cassette (fi g.2).
4. Add 1 additional drop of buffer diluent into the sample win-dow S if there is no beginning LF visible within 1 minute after adding the buffer diluent.

READING OF THE TEST RESULT

Read the test result 10 minutes after the buffer solution has been added into the sample win-dow S.

POSITIVE Anaplasma and / or Ehrlichia TEST RESULT (fi g.3–5)
A pink-purple TEST line of any intensity (varying from very weak to strongly intensive) and a pink-purple CONTROL line appear.

NEGATIVE TEST RESULT (fi g.4–6)
Only a pink-purple CONTROL line appears. This line indi-cates, irrespective of its intensity, that the test has been per-formed properly.

INVALID TEST RESULT
No CONTROL line visible. The test should be repeated using a new twin 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 twin test cassette to ensure a precise assignment.
• Use a new pipette and a new twin 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 infec-tious and disposed accordingly, together with the used test-kit components.

TEST PRINCIPLE

The FASTest® ANAPLASMA-EHRLICHIA is based on an immunochromatographic “sandwich principle”.
The antibodies against Anaplasma spp. and / or Ehrlichia canis present in the sample react in the conjugate pad with mobile antibodies, which are conjugated to colloidal gold particles. These antibody complexes are migrating along the nitrocellulose membrane (“lateral fl ow”, LF) and bind to fi xed recombinant Anaplasma spp. or Ehrlichia spp. anti-gens, forming 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 (stay abroad, tick infestation) and clinical data as well as therapy and prophylaxis possibili-ties.
• Any non-described colour or contour variation of T and C (e. g. greyish, shadow-like lines) has to be considered as unspecifi c reaction and therefore as negative test result.
• T can vary both in intensity (from weak to strong pink-purple) and in width. Therefore, any pink-purple line appearing within the required incubation time has to be interpreted as positive test result.
• Positive test results may be observed even before the end of incubation. Beyond this time, test results should not be interpreted.
• Due to anticoagulated whole blood and / or red hemo-globin background of the test membrane caused by hemolytic blood samples, the visibility of T could be from worse to not visible.
• For the detection of antibodies, a two-step diagnostics is known to be standard. The fi rst step starts with in-clinic IgG antibody screening test like FASTest® ANAPLASMA-EHRLICHIA. The suspicion about an active anaplasmosis /ehrlichiosis is substantiated by combination with ac-cording clinic. Furthermore, a quantitative antibody test-ing via IFAT (coupled serum samples at intervals of 2–4 weeks) should be taken to determine the end titre or the titre increase (seroconversion).

Positive test result

• The proof of ab against Anaplasma spp. / Ehrlichia canis, together with anamnesis and clinic, shows with a high likelihood that Anaplasma spp. / Ehrlichia canis can be considered as cause of the acute disease.
• Dog has had contact with Anaplasma spp. / Ehrlichia canis (ab formation!)
• Ab can persist over months to years inspite of therapy (potential chronic carriers).
• Asymptomatic, but anti-Anaplasma spp. / Ehrlichia canis ab positive animals have been infected with Anaplasma spp. / Ehrlichia canis at a particular time. Therefore, they are potential carriers of Anaplasma spp. / Ehrlichia canis. Subclinical animals could develop clinical symptoms at some indefi nite future date, especially in case of coinfec-tion with another vector-transmitted agent.
• The decision starting an antibiotic therapy should be based on indirect immunofl uorescence test (coupled serum test in an interval of 2–3 weeks with 3-fold titre increase) combined with clinical symptoms. Persistence of abs could be stable over a long term.

Negative test result

• With high likeliness, dog had no contact with Anaplasma spp. / Ehrlichia canis.
• Early stage of an Anaplasma spp. / Ehrlichia canis infec-tion (< 2–3 weeks post infection!). Dog has not formed abs in detectable concentration. Animals could still be seronegative during acute infection (ab increase from 14 days post-infection) resulting in a negative test result.

References

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

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