Standardized method and kit for the quantification of...

Details Standardized method and kit for the quantification of... Standardized method and kit for the quantification of...

C - Chemistry, Metallurgy

12

Q

C12Q 1/70 (2006.01)

Patent

CA 2689105

The present invention provides a standardized method and a kit for an accurate quantification of HAV in clinical and food samples. The general approach is based on the use of several controls to measure the efficiency of those critical steps of the quantification: the nucleic acids extraction and the RT-PCR reactions. The kit comprises: a Mengo virus mutant strain with the same growth properties than those of the wild-type Mengo virus and with no pathogenic capacity; a single stranded RNA molecule corresponding to a fragment of the HAV genome; primers that specifically bind to regions of the 5' non coding region of the HAV genome; a detectable labeled probe that specifically binds to the amplimer resulting from the RT-PCR; and an appropriate molecule to generate an standard curve for the quantification of HAV.

La présente invention concerne un procédé standardisé et une trousse utiles pour la quantification juste du VHA dans des échantillons d'aliments et des échantillons cliniques. L'approche globale repose sur l'utilisation de divers témoins pour mesurer l'efficacité des étapes critiques de la quantification de l'extraction des acides nucléiques et des réactions RT-PCR. La trousse comprend: une souche mutante de virus Mengo ayant les mêmes propriétés de développement que celles du virus Mengo de type sauvage et n'ayant pas de pouvoir pathogène; une molécule d'ARN simple brin correspondant à un fragment du génome VHA; des amorces qui se lient spécifiquement à des régions de la région non codante 5' du génome VHA; une sonde marquée détectable qui se lie de manière spécifique à l'amplimère résultant de la RT-PCR; et une molécule appropriée pour générer une courbe standard pour la quantification du VHA.

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