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发明授权
CN101027408B 血细胞抗原基因分型的方法和用于血细胞抗原基因分型的试剂盒 失效 - 权利终止

血细胞抗原基因分型的方法和用于血细胞抗原基因分型的试剂盒
摘要：
本发明提供了一种血细胞抗原基因分型的方法，其包括对来自一哺乳动物物种的个体的DNA进行多重聚合酶链反应(PCR)以便扩增并以可检测方式标记至少两种不同的血细胞抗原的基因座的含有血细胞抗原的核苷酸多态性位点的区域，并用如此扩增并标记的DNA片段来确定每一种所述血细胞抗原的基因型。多重PCR包括使用针对每一种待进行基因分型的血细胞抗原的至少一对血细胞抗原特异性嵌合引物和至少一种以可检测方式标记的通用引物，优选地为一对以可检测方式标记的通用引物。通用引物具有不存在于所述哺乳动物物种的DNA中的独特序列。每一嵌合引物对包括左侧嵌合引物和右侧嵌合引物，各引物包括3′端的血细胞抗原特异性部分和5′端的通用部分。嵌合引物的通用部分的碱基序列对应于所述至少一种通用引物的碱基序列。嵌合引物对的血细胞抗原特异性部分包含所述血细胞抗原的基因座的含有血细胞抗原的核苷酸多态性位点的区域。本发明还提供了用于通过此方法进行血细胞抗原基因分型的试剂盒，一组血细胞抗原特异性嵌合引物对和一组血细胞抗原等位基因特异性寡核苷酸探针。
摘要（英）：

A method of genotyping blood cell antigens comprising subjecting DNA from an individual of a mammalian species to a multiplex Polymerase Chain Reaction (PCR) to amplify and detectably label a region of the locus of at least two different blood cell antigens which contains the site of nucleotide polymorphism of said blood cell antigen and using the thus amplified and labeled DNA fragments to determine the genotype for each of said blood cell antigens. The multiplex PCR comprises the use of at least one pair of blood cell antigen-specific chimeric primers for each blood cell antigen to be genotyped and at least one detectably labeled universal primer, preferably a pair of detectably labeled universal primers. The universal primer(s) have a unique sequence not occurring in the DNA of said mammalian species. Each chimeric primer pair comprises a left chimeric primer and a right chimeric primer, each of them comprising a blood cell antigen-specific part at the 3' end and a universal part at the 5' end. The base sequence of the universal part of the chimeric primers corresponds to the base sequence of said at lea st one universal primer. The blood cell antigen-specific parts of the chimeric primer pair enclose a region of the locus of the blood cell antigen which contains the site of nucleotide polymorphism of said blood cell antigen. A k it for genotyping blood cell antigens by this method. A set of blood cell antig en- specific chimeric primer pairs and a set of blood cell antigen allele-specif ic oligonucleotide probes.

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