Vetlab Supplies FASTest BOR in TICK Test Kit Instructions

Vetlab Supplies FASTest BOR in TICK Test Kit

INFORMATION ON THE TEST-KIT

TEST-KIT COMPONENTS

1 test-kit FASTest® LEPTOSPIRA IgM contains:

• 1 or 5 test cassettes coated with monoclonal antibodies
• 1 dropper bottle A with 1.0 ml or 1.5 ml buffer diluent
• 1 or 5 sample tubes with squeezer
• 1 or 5 disposable plastic pipettes
• 1 tick remover
• 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.

INTRODUCTION

Leptospirosis (Weil’s disease) is a world-wide spread bacterial infectious disease in various animals and humans (zoonosis!). Of the pathogen species Leptospira interrogans sensulato, more than 300 serovars are known which are summarized in 24 serogroups of varying pathogenicity. Due to non-existence of a cross immunity of the vaccination serovars and an increasing “serovar shift”, leptospirosis becomes more important (world-wide increasing prevalences especially for L. icterohaemorrhagia, L. canicola, L. grippotypho- sa, L. australis and L. pomona).
Transmission is direct: horizontal (esp. infectious abortion material, urine, food animals [pathogen reservoir: rodents, small mammals], bites), vertical (lactogenic, placental, veneric) and indirect: contaminated soil, water.
Incubation time as well as symptoms are strongly depending on age, immune status, serovar type. Subclinical infections are rather the rule (higher seroprevalence than the prevalence of the clinical disease), proven by numerous studies. With present immunity due to past infection, normally quick antibody (ab) formation and pathogen elimination takes place. Typical are general symptoms like fever, apathy, anorexia, power drop, loss of weight, partially diarrhoea 3–7 days after infection as well as pale icteric mucous membranes. Other symptoms are late abortuses, dead births, birth of weak young animals. Within 48–72 h, a serious, partly deathly process can develop, depending on organ manifestation (especially kidneys [tubular persistence], liver and lung [hemorrhagic syndrome] dysfunctions, DIC).
A clinical suspicion for leptospirosis requires a quick, laboratory-ensured diagnosis, because the animals become a shedder and infection risk for humans and animals. Because the direct proof of the pathogen (dark field microscopy, culture, PCR [false negative through high antibiotic dose]) often is difficult, time consuming, expensive and only proving if positive, the ab detection, especially the IgM detection, has an important diagnostic relevance.
The actual reference method is microagglutination test (MAT). However, its sensitivity varies strongly (30–80 %) depending on the stadium of infection, and it does not distinguish between IgM and / or IgG antibodies. Therefore, a straight IgM detection (increase in the 1st week p. inf. / maximum from week 2–3 on), can be of significant diagnostic benefit compared to MAT or straight IgG detection (detectable not until 3–4 weeks, persistence for months, vaccination-caused IgG antibody persistence).
The FASTest® LEPTOSPIRA IgM is an important diagnostic tool for the veterinarian for the fast and simple on-site detection of a leptospirosis in the early stage of infection. Therefore, further laboratory diagnostics as well as therapeutic and prophylactic measures can be started immediately.

INFORMATION ON THE SPECIMEN MATERIAL

10 µl (defined volume of attached plastic pipette) of 15–25 °C warm plasma (P), whole blood supernatant (WBS: supernatant from whole blood, with anticoagulant, without centrifugation), or serum (S) are needed. Mix the sample material well before use!
Non-cooled (15–25 °C), P, WBS and S should be tested within 4 hours! At 2–8 °C, P, WBS and S can be stored up to 4 days. The 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, fibrinogen, 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 influence the target measurement. These can lead to an impaired LF and / or unspecific reactions on T and C.
Whole blood should not be used in the FASTest® LEPTOSPIRA IgM.

SPECIMEN PREPARATION (Small and large tick)

• a. Open the sample tube and place the tick in the middle of the sample tube (fi g.1).
• b. Squeeze the tick by closing the blue cap with the attached squeezer tightly onto the sample tube (fi g.2).

Small tick (Ø below 5 mm)

• c. Open the sample tube again. Hold the dropper bottle A vertically and add 4 drops of buffer diluent (ca. 160–200 µl). In case of any tick remnants on the squeezer, let the 4 drops fl ow over the squeezer into the sample tube (fi g.3a).
• d. Mix the squeezed tick material homogeneously with the buffer diluent by twisting and untwisting the blue cap for several times (see fi g.2).

Large tick (Ø above 5 mm)

• c. No addition of buffer diluent!

TEST PROCEDURE

1. Remove the test cassette from its foil pouch shortly be-fore use. Place it on a flat surface.
   Small tick (Ø below 5 mm)

2. Suck up the whole tick- buffer mixture (TBM, avoid tick particles if possible) using the disposable plastic pipette (fi g.4a). Add 3 drops (120–150 µl) slowly (without air bubbles, one drop after the other) into the sample window S of the test cassette (hold pipette vertically, fi g.5a). Rest volume, see chapter 9.

Large tick (Ø above 5 mm)

• Suck up the whole tick mixture (TM, avoid tick particles if possible) using the disposable plastic pipette (fi g.4b). Add 1 drop (40–50 µl) into the sample window S of the test cassette (hold pipette vertically, fi g.5b). Rest volume, see chapter 9.
• Add 2 drops (80–100 µl) buffer diluent into the sample window S of the test cassette (fi g.6).

If there is no beginning LF visible within 1 minute after adding the TBM (5.2) or the buffer (5.3), mix the TBM / the buffer in the sample window S with light pressure of the used pipette tip onto the membrane or add 1 drop of buffer diluent.

Large tick (Ø above 5 mm)

READING OF THE TEST RESULT

Read the test result 15 minutes after the buffer diluent has been added into the sample window S.

POSITIVE TEST RESULT (fi g.7)

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.8)

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 (protective 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 disposable plastic pipette and a new test cassette for each sample.
• The buffer diluent contains 0.1 % ProClin™ 950 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® LEPTOSPIRA IgM is based on an immuno-chromatographic “sandwich principle”.
In the conjugate pad, the sample IgM antibodies against Leptospira will react to mobile monoclonal anti-dog anti-bodies which are bound to colloidal gold particles. Migrating (“lateral fl ow”, LF) along the nitrocellulose membrane, these specific antigen-antibody complexes are bound by fixed Leptospira 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 color or contour variation of T and C (e. g. greyish, shadow-like lines) has to be considered as unspecific reaction and therefore as negative test result.
• Positive test results may be observed earlier, depending on the concentration of antigen in the sample.
• Due to red hemoglobin background of the test membrane, caused by bloody sample material (large totally soaked up ticks), the visibility of T, especially in case of weak positive samples, could be from worse to not visible.
• T can vary both in intensity (from weak to strong pink-purple) and in width. Therefore, any pink-purple line which appears within the required incubation time has to be interpreted as a positive test result.
• Rest volume (TBM or TM) can be used for PCR confirmation.

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

References

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

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