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PCR Test for the Detection of Neosporosis


418023

Código: 418023. Categorías: , , .
   
Descripción

1. Purpose of the test
ADIAVET™ NEOSPORA REAL TIME kit is intended to detect Neospora caninum using real-time
Polymerase Chain Reaction (PCR) technology from swab, from tissue and brain specimens of bovine,
ovine and equine.
2. Pathogen
Neospora caninum is an apicomplexan parasite first associated with bovine abortion in the USA in 1989
(Bjerkas et al., 1984; Thilsted and Dubey, 1989). Since this observation N. caninum has been implicated
as one major cause of infectious abortion of cattle worldwide.
Abortion due to neosporosis can occur at any stage of pregnancy but is most likely to occur between 5–
7 months of gestation. Cows that aborted in a previous pregnancy due to neosporosis can abort again.
They can also give birth to diseased calves, calves with a sub-clinical N. caninum infection, or
uninfected calves.
Neospora caninum is a cyst-forming coccidian parasite. Two asexual stages of development occur in
the intermediate host (cattle, horse), tachyzoite and bradyzoite and one sexual stage (oocyst) occurs in
the definitive host (dog) (McAllister et al ., 1998).
Routine diagnosis of N. caninum infection is made by observation of clinical signs (abortion, peaking
between the fifth and seventh month of gestation), by histology of foetal tissue and by indirect
fluorescent antibody (IFAT), and ELISA tests on maternal and foetal blood. But abortion is not always
related to the presence of antibody.
In 1996, a shift of biologist of Institute of Parasitology of Berne conducted by the Professor Kaufmann
identified a repetitive genomic fragment called Nc5, specific of N. caninum (Kaufmann et al ., 1996).
Since, the sequence has been used as probe in molecular test as PCR test (Muller et al., 1996; Yamage
et al., 1996).
The PCR test allows the specific and sensitive detection of the parasite from foetal tissues.
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™ NEOSPORA REAL TIME kit enables the simultaneous detection of:
– N. caninum (probe labelled in FAM),
– GAPDH, an internal control of extraction and amplification steps specific from an
endogenous DNA (probe labelled with a fluorochrome read in the same spectra as VIC
and HEX).
ADIAGENE recommands the test using DNA purification kits (Qiagen and Macherey-Nagel). Other
purification kits can be used if they have been validated by the user.

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