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PCR Test for the Detection of Swine Flu


ADI282-100

Código: ADI282-100. Categorías: , , .
   
Descripción

1. Purpose of the test
ADIAVET™ SIV REAL TIME kit is intended to detect the Swine Influenza Viruses (SIV) using real-time
Polymerase Chain Reaction (PCR) technology from nasal swab, allantoic or bronchial fluid and tissue
specimens of pig, as well as from viral culture.
2. Pathogen
The swine flu is an important pig farm disease because it could lead to severe respiratory symptoms. It is
generally characterized by a high number of infected animals but a low mortality level. The disease is
due to a virus of the Influenza type A genus. The first isolation of an influenza virus on pig occurs in
1930 (Influenza type A, H1N1). In opposition to the human flu, Swine flu could spread in pig farms all
over the year. Three sub-types of Influenza type A virus can usually infect pigs: H1N1, H3N2 and H1N2.
H1N1 and H1N2 are enzootic in Europe.
The detection of the virus is usually performed by viral culture on nasal swabs. Viral culture as well as
diagnostic tests based on the detection of nucleoprotein antigen (such as immunofluorescence staining
assay) can be carried out on lung sample. Viral culture has to be performed at an early stage of the
disease because the virus persists only few days at the lesions level.
Real time PCR can be an interesting, specific and sensitive alternative method to obtain a result in one
day if nasal swabs are taken on the early first clinical signs (exhaustion, fever, loss of appetite,
respiratory difficulties).
3. Description and purpose of the test
This test is based first on the reverse transcription (RT) of RNA into complementary DNA (cDNA). 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™ SIV REAL TIME kit enables the simultaneous detection of:
– swine influenza virus (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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