Primer-specific and mispair extension assay for identifying...

C - Chemistry – Metallurgy – 12 – Q

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C12Q 1/68 (2006.01)

Patent

CA 2339323

The invention relates to primer-specific and mispair extension assays for identifying gene variations, such as in different genotypes or subtypes of a given genotype. The assay includes extending a nucleic acid sequence from a patient sample with extension products of the primer, characterizing the extension products, and comparing the extension products with known nucleic acid sequences of various genotypes for determining the genotype of the nucleic acid sequence extended. The one or more extension products are terminated in the presence of at least two mispairs within a 2 to 4 base pair range located downstream of the 3' end of the primer. In the assay, at least one primer or the dNTPs is labeled.

L'invention concerne une méthode d'extension de mésappariement à spécificité d'amorçage servant à identifier des variations de gènes, telles que des variations dans différents génotypes ou sous-types d'un génotype donné. Cette méthode consiste à (a) effectuer l'extension d'une séquence d'acides nucléiques provenant d'un spécimen prélevé sur un patient par l'intermédiaire d'une polymérase au moyen d'une amorce spécifique pour un génotype à déterminer et d'un ensemble incomplet de dNTPs dans des conditions permettant d'obtenir des produits d'extension de l'amorce ; (b) caractériser ces produits d'extension ; (c) comparer ces produits d'extension à des séquences d'acides nucléiques connues de différents génotypes afin de déterminer le génotype de la séquence d'acides nucléiques soumise à l'extension. Dans cette méthode, au moins un de l'amorce ou des dNTPs est marqué.

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