公开(公告)号：EP1730311B1

公开(公告)日：2010-09-15

申请号：EP05729402.7

申请日：2005-03-31

申请人： Stichting Sanquin Bloedvoorziening

发明人： BEIBOER, Sigrid, Herma, Wilma ,  WIERINGA-JELSMA, Hendrika ,  DEN DUNNEN, Johannes, Theodorus ,  DE HAAS, Maschenka

IPC分类号： C12Q1/68

CPC分类号： C12Q1/6827 ,  C12Q1/686

摘要： 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 least 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 kit for genotyping blood cell antigens by this method. A set of blood cell antigen-specific chimeric primer pairs and a set of blood cell antigen allele-specific oligonucleotide probes.

公开(公告)号：EP1730311A1

公开(公告)日：2006-12-13

申请号：EP05729402.7

申请日：2005-03-31

申请人： Stichting Sanquin Bloedvoorziening

发明人： BEIBOER, Sigrid, Herma, Wilma ,  WIERINGA-JELSMA, Hendrika ,  DEN DUNNEN, Johannes, Theodorus ,  DE HAAS, Maschenka

IPC分类号： C12Q1/68

CPC分类号： C12Q1/6827 ,  C12Q1/686

摘要： 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 least 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 kit for genotyping blood cell antigens by this method. A set of blood cell antigen-specific chimeric primer pairs and a set of blood cell antigen allele-specific oligonucleotide probes.