Process for typing hla-b using specific primers and probes sets

Details Process for typing hla-b using specific primers and probes sets Process for typing hla-b using specific primers and probes sets

C - Chemistry, Metallurgy

12

N

C12N 15/12 (2006.01) C07H 21/00 (2006.01) C12P 19/34 (2006.01) C12Q 1/68 (2006.01) C40B 40/06 (2006.01)

Patent

CA 2158578

The invention relates to a method for typing or subtyping one or more HLA-B alleles characterized by the sequence GCCA at position 30 to 33 of exon 2 (with said numbering being according to Zemmour and Parham, 1992), liable to be present in a sample, with said method comprising at least the following steps: (i) amplifying HLA-B alleles with at least one 5' end amplification primer selected from the following list: 5' -AGGTATTTCTACCCGCCA-3' (B25P) or sequence variants thereof, in combination with an appropriate 3' end primer being chosen from the same alleles as the above defined 5' end primers, with said 5' and 3' end primers being possibly labelled; and, (ii) hybridizing the amplified product, being labelled during or after amplification, at appropriate conditions with one or more suitable probes selected from region 15 to 261 of the HLA-B exon 2 region, with said numbering being according to Zemmour and Parham, 1992, (iii) washing at appropriate washing conditions, (iv) detecting the hybrids formed; and, (v) inferring the allele present from the observed hybridization pattern.

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