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PCR Test for the Detection of Besnoitiosis


Código: ADI451-100. Categorías: , , .
   
Descripción

1. Purpose of the test
ADIAVET™ BESNOITIA REAL TIME kit is intended to detect Besnoitia besnoiti using real-time
Polymerase Chain Reaction (PCR) technology from skin and whole blood specimens of bovine.
2. Pathogen
Besnoitia besnoiti, the causative agent of bovine besnoitiosis, is an obligate intracellular protozoan. The
disease affects mainly young cattle. Besnoitiosis is epizootic in the south of France, but is now widely
distributed in Africa, Asia and in Southwestern Europe. The most likely pathway of transmission would
be transcutaneous, by stinging insects (tabanids, stomox).
During infection, an incubation stage of 3 to 6 days is followed by 3 successive clinical stages:
– A febrile stage of 3 to 7 days; the tachyzoites multiplication in endothelial cells of blood vessels
increases the animal temprerature
– A second stage of 1 to 2 weeks; the bradyzoites cysts generate subcutaneous oedema
– A chronic stage of several months, characterised by alopecia and sclerodermia. The skin becomes
then markedly thickened and wrinkled, and parasitic cysts are observed on conjunctiva and sclera.
This ultimate phase leads generally to the death of the animal or to its euthanasia.
Serologic tests are available for the detection of the specific antibodies of Besnoitia besnoiti present in
the chronic stage. In order to avoid the transfers contaminated animals and to control the spread of the
bovine besnoitiosis, it is essential to use diagnostic tools that detect the pathogens at the early stages
of the disease.
PCR tests are able to detect the parasites by the detection of tachyzoites of Besnoitia besnoiti during
febrile stage in the monocytes of blood and in the skin during oedema and chronic stages.
3. Description and purpose of the test
This test is based on enzymatic gene amplification or PCR technology.
Amplified products are detected in real-time thanks to a specific labelled hydrolysis probe (5’-
exonulease technology).
The ADIAVET™ BESNOITIA REAL TIME kit enables the simultaneous detection of:
– Besnoitia besnoiti (probe labelled in FAM),
– GAPDH, an internal control of extraction and amplification steps specific from an
endogenous DNA (probe labelled with a fluorochrome with the same spectra as VIC or
HEX).
ADIAGENE validated the test using DNA purification kits (Qiagen, Macherey-Nagel). Other
purification kits can be used if they have been validated by the user.

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