Genodive Pharma Inc.

Hidetoshi Inoko

Shingo Suzuki

Takashi Shiina

Yuki Wada

Shigeki Mitsunaga

The purpose of the present invention is to provide a method and kit for highly precise DNA typing, in which a high throughput sequencer is used and ambiguity derived from phase ambiguity is eliminated. The present invention provides a method for the DNA typing of HLA, which is characterized by comprising: (1) a step of preparing sets of primers which respectively hybridize specifically to an upstream region and a downstream region of at least 2 genes selected from genes belonging to HLA class I and HLA class II in a human genome sequence, and are capable of amplifying under the same PCR conditions; (2) a step of simultaneously amplifying said at least 2 genes in a test sample (DNA) using the sets of primers in a single container under the same PCR conditions; (3) a step of determining the nucleotide sequences of PCR amplified products; and (4) a step of optionally carrying out a homology search within a database.

Method and kit for multiplex DNA typing of HLA gene

The purpose of the present invention is to provide a method and kit for highly precise DNA typing, in which ambiguity derived from phase ambiguity is eliminated. The present invention provides a method for the DNA typing of HLA, which is characterized by comprising: (1) a step of preparing a set of primers which can respectively anneal specifically to an upstream region and a downstream region of each of HLA-A, HLA-B, HLA-C, HLA-DQA1, HLA-DQB1, HLA-DPA1 and HLA-DPB1 gene in the nucleotide sequence for the human genome, and a set of primers which can respectively anneal specifically to exon-2 and a 3'-side non-translated region in HLA-DRB1; (2) a step of carrying out the PCR amplification of a sample to be tested (DNA) using the sets of primers; (3) a step of determining the nucleotide sequence for a PCR-amplified product; and (4) an optional step of carrying out the homology search in a data base.

Method and kit for DNA typing of HLA gene

The present invention addresses the problem of providing a method and kit for the DNA profiling of HLA genes using a high-throughput massively parallel sequencer. The present invention pertains to a method for the DNA profiling of HLA genes, said method being characterized by including: (1) a step for preparing a primer set that anneals specifically to exon 4 and intron 1 and includes exon 2 and exon 3 of at least one target gene selected from the group consisting of HLA-DRB1, HLA-DRB3, HLA-DRB4, HLA-DRB5, HLA-DQB1 and HLA-DPB1 in the base sequence of the human genome; (2) a step for amplifying a sample (DNA) by PCR using the primer set; (3) a step for determining the base sequence of the amplified PCR product; and (4) a step for carrying out a homology search against a database.

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Kanagawa, Japan

Hidetoshi Inoko