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PCR Test for the Detection of Avian Flu


ADI283-100

Código: ADI283-100. Categorías: , , .
   
Descripción

1. Purpose of the test
ADIAVET™ AIV REAL TIME kit is intended to detect Avian Influenza Viruses (AIV) using real-time
Polymerase Chain Reaction (PCR) technology from swab, tissue, faeces specimens of poultry and viral
strain culture.
2. Pathogen
All Avian influenza (AI) viruses belong to the Influenzavirus A genus of the orthomyxoviridae family.
They are negative single stranded RNA viruses divided into subtypes based on two surface proteins:
hemagglutinin and neuraminidase. AIV can cause severe diseases in domestic poultry, including
chickens and turkeys but can also infect pheasants, quails, ducks, geese…
AIV is spread through direct contact (bird to bird) but also through indirect contact with contaminant
equipment and materials. The virus is excreted through infected birds faeces and secretions from the
respiratory tract.
Currently, virus isolation in embryonating chicken eggs and subtyping by serological methods
constitute the standard for AIV detection, but it may take 1 to 2 weeks.
Real-time PCR could be an alternative method to obtain result within one day, with a high specificity
and sensitivity.
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™ AIV REAL TIME kit enables the simultaneous detection of:
– Avian 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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