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PCR Test for the Detection of Classical Swine Fever


ADI222-100

Código: ADI222-100. Categorías: , , .
   
Descripción

1. Purpose of the test
ADIAVET™ CSFV REAL TIME kit is intended to detect the Classical Swine Fever Virus (CSFV) using realtime
Polymerase Chain Reaction (PCR) technology from tissue, whole blood and serum specimens of
pig and wild boar, as well as from viral culture.
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). CSFV is listed in the A OIE list; it causes
heavy epizooties by pigs, and so presents a risk for environment. Generally, all precaution measures
have to be taken to avoid viral particles to spread out of laboratory. Refer to the reference centre of
your country to know the requirements for biosecurity (experiments in a certified laboratory).
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™ CSFV REAL TIME kit enables the simultaneous detection of:
– CSFV (probe labelled in FAM),
– GAPDH, 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
and HEX).
ADIAGENE validated the test using RNA purification kits (Qiagen, Macherey-Nagel). Other purification
kits can be used if they have been validated by the user.

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