Methods and reagents for typing hla class i genes

C - Chemistry – Metallurgy – 12 – Q

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C12Q 1/68 (2006.01) C07H 21/04 (2006.01) C07K 14/74 (2006.01)

Patent

CA 2213731

A method for typing HLA class 1 genes. The method entails first contacting a sample DNA with first and second amplification primers, wherein the first and second primers are each at least partially located in an exonic region. Next, using the first and second primers, a target sequence is amplified by the PCR to form an amplicon of the target sequence. Finally, the amplicon is detected with a sequence-specific detection means, e.g., DNA sequencing. The invention also includes specific amplification primers, specific sequencing primers, and kits especially adapted for use with the above HLA typing method.

La présente invention concerne un procédé de typage de gènes HLA de classe I. Ce procédé consiste à mettre un échantillon d'ADN en présence d'une première et d'une seconde amorce d'amplification au moins partiellement situées dans une région exonique. Le procédé consiste ensuite, en utilisant la première et la seconde amorce, à amplifier la séquence par amplification en chaîne de la polymérase de façon à former un amplicon de la séquence cible. Le procédé consiste enfin à détecter l'amplicon au moyen d'un outil de détection spécifique de la séquence, par exemple, un séquençage ADN. L'invention concerne également des amorces spécifiques de l'amplification, des amorces spécifiques du séquençage, et des trousses d'analyse spécialement conçues pour le procédé de typage HLA de l'invention.

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