The rapid slide agglutination test (RSAT) for the diagnosis of Brucella canis was described by George and Carmichael (Proc. Council Res. Workers in Animal Disease, November 1973). The basis of the Genprice Gentaur test is direct agglutination of the killed stained antigen by Brucella canis antibodies.
GENERAL INFORMATION
- Canine brucellosis is a chronic infection with Brucella canis that causes generalized lymphadenitis and mild to severe reproductive symptoms. Prostatitis, epididymitis, scrotal dermatitis, testicular atrophy and impotence can be present in the male; abortion typically in the final two weeks of gestation, resorption, infertility, and vaginal discharge can be present in the female.
- Brucella canis is infectious to humans. Caution should be exercised when handling serums to be tested.
- The test has been shown to presumptively diagnose infection with Brucella canis by detecting specific antibody that is formed 1 to 4 weeks after infection. The basis for serodiagnosis of canine brucellosis is the 2-Mercaptoethanol Tube Agglutionation Test (2ME-TAT) and the Rapid Slide Agglutination Test (RSAT).
- The 2ME-TAT and RSAT have demonstrated excellent correlation in experimentally infected dogs. In field situations, it has been recognized that an occasional healthy dog, culturally negative for
- Brucella canis will react positively in the RSAT, but not in the 2ME-TAT. The 2-Mercaptoethanol – Rapid Slide Agglutination Test (2ME-RSAT) has been developed in an attempt to eliminate discrepancies between the RSAT and the 2-ME-TAT.
- Certain non-specific agglutinins, reported to occur in the sera of normal dogs, are removed from canine sera when 2-mercaptoethanol is employed in the 2ME-TAT. Because of this occurrence, 2-
mercaptoethanol is employed in the modified Rapid Slide Agglutination Test procedure.
INDICATIONS
The reagents are used to presumptively diagnose infection with Brucella canis in dogs. Positive results
require additional confirmation.
PRECAUTIONS
Antigen and accompanying antiserum have been standardized and should be used together. Store
components at 2-7C (35-45F). Do not allow reagents to stand at room temperature for excessive
periods of time. Do not freeze.
SAMPLE INFORMATION
Serum is required. Test may not be used with whole blood or plasma.
Procedure for Rapid Slide Agglutination Test (RSAT)
1. ENSURE REAGENTS AND SAMPLE ARE AT ROOM TEMPERATURE (70-78F; 21-25C) prior
to testing.
2. Cut card on dotted line. Two circles are used to perform each test.
3. Place one drop of Reagent A, Positive Control (Red Cap), in one circle of the supplied card.
4. Use a disposable plastic pipette to place one drop of test serum in another circle. Each test kit
contains disposable pipettes to prevent serum cross contamination. Do not dispose of pipette until
the 2ME-RSAT is completed.
5. Thoroughly mix Reagent B, Antigen (White Cap), by vigorously shaking the vial. Add one drop
of the Reagent B, Antigen, close to each serum drop, being careful not to touch the serum with
the dropper tip.
6. Mix each antigen-serum combination with a separate end of a stir stick, spreading to fill the circular
area. Do not allow the positive control test to touch the unknown serum sample.
7. Gently rock the card for 10-15 seconds. Place on a flat surface and observe for agglutination for
no longer than two minutes. If the serum is negative (absence of agglutination), the animal is
considered not to be infected with B. canis. If the card tests positive, perform the 2ME-RSAT.
Procedure For 2-Mercaptoethanol-Rapid Slide Agglutination Test (2ME-RSAT)
1. Add 2 drops of Reagent C, 2-Mercaptoethanol (Yellow Cap) to a tube containing 2 drops of the
serum to be tested, and mix well.
2. Place 1 drop of this mixture on a new reaction circle.
3. Add 1 drop of Reagent B, Antigen as above to the serum solution and mix with stir stick as
described above.
4. Gently rock the card for 10-15 seconds. Place on a flat surface and observe for agglutination for
no longer than 2 minutes. When the RSAT-positive sample also tests positive by 2ME-RSAT, the
animal is presumptively diagnosed as being infected with B. canis. Blood should be subjected to
cultural examination for B. canis. When the RSAT-positive samples tests negative by 2ME-RSAT,
the animal may be in the early stage of B. canis infection, or alternatively, its serum may contain
non-specific agglutinins to B. canis. To distinguish between these two conditions, a second serum
sample should be collected in approximately thirty days and retested by the 2ME-RSAT procedure.
Only if this sample tests positive should the animal be presumptively diagnosed as having B. canis
infection. Blood should then be subjected to cultural examination for B. canis.
Definitive diagnosis of canine brucellosis is based upon isolation of B. canis from the animal.
KIT CONTENTS
10 Tests/Kit 25 tests/Kit
Reagents A – Positive Control 1 vial 1 vial
Reagent B – Antigen 1 vial 1 vial
Reagent C – 2-Mercaptoethanol, (0.2M solution) 1 vial 1 vial
Reaction cards 5 each 13 each
Pipettes 10 each 25 each
Stir Sticks 10 each 25 each
Background
Brucellosis, also known as undulant fever, Mediterranean fever is a zoonosis and the infection is almost invariably transmitted by direct or indirect contact with infected animals or their products. Expansion of animal industries and urbanization, and the lack of hygienic measures in animal husbandry and in food handling partly account for brucellosis remaining a public health hazard. Expansion of international travel which stimulates the taste for exotic dairy goods such as fresh cheeses which may be contaminated, and the importation of such foods into Brucella-free regions, also contribute to the ever-increasing concern over human brucellosis.
Introduction
The kit is based on sandwich immunochromatographic assay, to detect bovine brucellosis antibody in serum / plasma / blood, which is rapid, accurate and easy-to-operate.
Qualitative Detection Of IgG Antibodies Against Brucella Canis
Sample: Whole blood, Plasma or SerumFASTest Brucella Canis is a rapid immunochromatographic test for the detection of IgG antibodies in the anti-coagulated whole blood, plasma or serum of dogs. Showing high Sensitivity of 90.5% and Specificity of 88.4%, Brucella Canis test kit enables the veterinarian to confirm quickly and on-site an estimated diagnosis and to start therapy and a Brucellosis Prevention Program adapted to the breeder’s needs.
Abortions and Reproductive Failures
Brucellosis in the dog shows low prevalence but occurs most commonly after ingestion of contaminated placental materials or aborted foetuses, vaginal discharges from infected bitches that are in heat or who abort, and during breeding. Following an abortion, Brucella Canis may be shed for several weeks or, intermittently, for months.
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Brucella Canis is a zoonotic organism and it is different for male and female dogs. Treatment for the disease can often be very expensive and it is thought that a combination of minocycline and streptomycin is useful but unaffordable.