Vetlab Supplies FASTest NEOSPORA Caninum Test Kit Instructions

Vetlab Supplies FASTest NEOSPORA Caninum Test Kit

INFORMATION ON THE TEST-KIT

TEST-KIT COMPONENTS

1. test-kit FASTest® NEOSPORA caninum contains:
2. 2 or 10 test cassettes coated with recombinant Neospora caninum antigens
3. dropper bottle A with 1.0 ml or 3.0 ml buffer diluent
4.  or 10 disposable plastic pipettes
5.  instructions for use

STABILITY AND STORAGE

APPLICATION AND ABBREVIATIONS

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 96.2 %
• Specifi city 100 %
• (Comparison Method: IFAT)

INTRODUCTION

Neospora caninum plays an important epidemiological role in dogs and cattle. The dog (esp. watch dogs, stray dogs) is a defi nitive host (intestinal neosporosis) and excretes infectious oocysts with the feces. The intermediate host, especially cattle, but also goats, sheep and horse, gets infected via grazing land and / or water contaminated with oocysts (horizontal infection) and / or intrauterine via already infected mothers (vertical infec-tion). N. caninum plays an important role in abortion in cattle world-wide. Characteristics are accumulating abortions in all states of gestation, dead births and weak calves. The dog can act as an intermediate host as well and therefore can fall sick with neosporosis (systemic neosporosis). In dogs, the symptoms are especially focused on neurological disorders: paresis / paralysis of the hind-limbs, later also of the fore-limbs, as well as polymyelitis, radiculitis and encephalomyelitis. Also, muscular atrophy, hyperextension, hyperaesthesia and dyspha-gia can occur. Additionally, hepatitis, pneumonitis, myocarditis and ulcerative dermatitis can appear. In older dogs Neospora infection usually is asymptomatic! Puppies become clinically conspicuous at the age from 3 to 9 weeks up to one year. Early diagnostics and therefore specifi c therapy are essential for the prognosis. Due to recent studies, there seems to be a predispo-sition of male dogs to N. caninum.

Dog: Due to the short excretion period and the low amount of oocysts in dog feces, the detection of antibodies using FAST-est® NEOSPORA caninum becomes very important for the diag-nosis of a neosporosis.

Cattle: Due to horizontal, but particularly to the economically more important vertical placental transmission of infection onto the offspring, with FASTest® NEOSPORA caninum suspicious stocks should be tested for N. caninum antibodies.

INFORMATION ON THE SPECIMEN MATERIAL

Approximately 50 µl (2 drops of attached plastic pipette) 15–25 °C warm whole blood (WB, with anticoagulant) or 25 µl (1 drop of attached plastic pipette) 15–25 °C warm plasma (P) or serum (S) are needed. Native blood without any anticoagulant must be avoided due to the potential risk of microclots (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. 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 temperature (15–25 °C) 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 B 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 be fore use. Place it on a fl at surface.
2. Add 2 drops of anticoagulated whole blood (ca. 50 µl) or 1 drop of plasma / serum (ca. 25 µl) with the attached plastic pipette into the sample window A of the test cas-sette (hold the pipette vertically, fi g.1).
3. Hold the dropper bottle A vertically and express 4 (four) drops (160–200 µl) of buffer diluent into the sample win-dow A of the test cassette (fi g.2).
4. Add 1 additional drop of buffer diluent into the sample window A 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 15 minutes after the four drops have beed added into the sample window A.

POSITIVE TEST RESULT (fi g.3)
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)
Only a pink-purple CONTROL line appears. This line in-dicates, 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.

• 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 en-sure a precise assignment.
• Use a new pipette and a new test cassette for each sam-ple.
• 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 of accordingly, together with the used test-kit components.

TEST PRINCIPLE

The FASTest® NEOSPORA caninum is based on an immu-nochromatographic “sandwich principle” technique detect-ing specifi c antibodies against Neospora caninum in the whole blood, plasma or serum of the dog or cattle. The antibodies against Neospora caninum present in the sample will react in the conjugate pad with mobile mono-clonal antibodies, which are conjugated to colloidal gold particles. These antibody complexes are migrating (“lateral fl ow”, LF) along the nitrocellulose membrane and bind to fi xed recombinant Neospora caninum antigens forming a pink-purple TEST line (B). 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 outline variation of B and C (e. g. greyish, shadow-like lines) has to be considered as unspecifi c reactions and therefore as negative test result.

• Due to anticoagulated whole blood and / or red hemo-globin background of the test membrane, caused by hemolytic blood samples, the visibility of B, especially in case of weak positive samples, could be from worse to not visible.

• The proof of anti-Neospora caninum antibodies, together with anamnesis and clinic shows with a high likelihood that N. caninum can be considered as cause of the acute disease.

• In asymptomatic animals and antibody positive animals, one should assume that they could have been infected with N. caninum and, as potential carriers, can transmit
  N. caninum vertically on their offspring.

• Additionally, the titre amount or the titre increase (within two tests in an interval of 2–3 weeks) can be defi ned via indirect immunofl uorescence. A proof of the pathogen can be done by PCR in foetal fl uids or tissue samples of deceased whelps or in the liquor / muscle biopsies of liv-ing animals.

• For humans, there is no zoonotic risk at all.

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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