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PCR Test for the Detection of Bovine Viral Diarrhea and Border Disease


Código: ADI105-100. Categorías: , , .
   
Descripción

1. Purpose of the test
ADIAVET™ BVDV REAL TIME kit is intended to detect the Bovine Viral Diarrhoea Virus (BVDV) and the
Border Disease Virus (BDV), using real-time Polymerase Chain Reaction (PCR) technology, from whole
blood, serum, and tissue specimens of bovine, ovine, caprine and wild cervid, from ear notch of bovine,
as well as from milk specimen of bovine, ovine and caprine.
2. Pathogen
Pestiviruses consist in a single strand of positive sense RNA. Bovine Viral Diarrhoea Virus (BVDV), classical
swine fever (CSFV) and border disease virus (BDV) in sheep are also members of the pestivirus genus
which belongs to the Flaviviridae family (like hepatitis C). BVDV, which induces mucosal disease in
bovine, causes economic losses in cattle.
Many countries have started eradication programs of this disease, which involves a perfect management
of infected animals. Indeed, those must be detected earlier with a high reliability. However, the prenatal
infection of a calf between the 60th and the 120th day of gestation leads to the birth of a persistently
infected (PI) animal. These contagious animals are seronegative all their live and positive by virology. The
detection of the virus by antigenemy is only possible several weeks after their birth because of the
persistence of colostral antibodies. The earlier detection of these persistently infected animals is still
necessary in eradication programs.
Since the discovery of DNA in vitro amplification in 1985 (PCR), many scientists have developed virus
screening tests using genomic amplification of the RNA also called RT-PCR. Most of these tests allow the
detection of minute quantities of BVDV in blood or organs of infected animals, even with less than three
months old animals.
3. Description and purpose of the test
This test is based first on the reverse transcription (RT) of RNA into complementary DNA. Then, cDNA is
amplified (PCR) by a DNA polymerase using specific primers. Both enzymatic reactions occur in the same
tube (One-step RT-PCR).
Amplified products are detected in real-time thanks to a specific labelled hydrolysis probe (5’-
exonulease technology).
The ADIAVET™ BVDV REAL TIME kit enables the simultaneous detection of:
– BVDV, BDV and CSFV (probe labelled in FAM),
– RNaseP, an internal control of extraction and amplification steps specific from an
endogenous RNA (probe labelled with a fluorochrome read in the same spectra as VIC or
HEX).
ADIAGENE recommends using this test with RNA purification kits (Adiagene, Qiagen or MachereyNagel).
Other purification kits can be used if they have been validated by the user.

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