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Utilisation de la Rt-PCR En Temps Réel
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ISBN10: 6203897175
ISBN13: 9786203897173
Publisher: Editions Notre Savoir
Published: Apr 24 2025
Pages: 68
Weight: 0.23
Height: 0.16 Width: 6.00 Depth: 9.00
Language: French
ISBN13: 9786203897173
Publisher: Editions Notre Savoir
Published: Apr 24 2025
Pages: 68
Weight: 0.23
Height: 0.16 Width: 6.00 Depth: 9.00
Language: French
Le virus de la fièvre aphteuse (FMDV) est diagnostiqué par un test d'isolement du virus combiné à un test ELISA de l'antigène et, depuis peu, par une RT-PCR en une seule étape. L'utilisation de la RT-PCR fluorogénique prend de plus en plus d'importance pour la détection rapide du virus de la fièvre aphteuse à partir d'échantillons cliniques qui ne peuvent pas être traités par des tests d'isolement du virus et des tests ELISA de l'antigène. La RT-PCR quantitative en temps réel (qRT-PCR) combine la transcription inverse avec l'amplification par PCR et la détection de l'amplification positive à l'aide de sondes fluorescentes telles que la sonde TaqMan (R). Dans la présente étude, une qRT-PCR basée sur la sonde TaqMan(R) a été standardisée et utilisée pour la détection du génome de la fièvre aphteuse à partir d'échantillons cliniques comprenant des sécrétions nasales, du plasma et des fluides oesophago-pharyngés (échantillons probang). La standardisation initiale a été réalisée en utilisant différentes dilutions en série d'un isolat du virus de la fièvre aphteuse OTNN 24/84 (106,8 DICT50) et 102 virus ont pu être identifiés à l'aide de la TaqMan(R) qRT-PCR. Des standards d'ARN ont été synthétisés in vitro à partir d'un plasmide contenant 110 paires de bases de la région 3D d'un isolat du virus de la fièvre aphteuse de type O. L'ARN a été dilué dix fois en série et utilisé comme ARN standard pour générer des courbes standard.
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