[发明专利]一种用于人白细胞抗原基因分型的液相基因芯片系统

摘要

本发明涉及生物医学领域，尤其涉及人白细胞抗原基因分型的检测方法和试剂。选择人白细胞抗原基因复合体中多态性最丰富的HLA－A、－B、－DRB1三个位点的主要变异区设计、合成一套探针，探针共价连接到荧光微球载体表面，每种不同荧光的微球耦联一种探针，结合不同探针的荧光微球等量混合，组成液相基因芯片。本发明实现了多个基因片段的均衡扩增，探针与目标基因片段杂交在液相环境中进行，相对于传统基因芯片的液－固相杂交，杂交效率更高。另外，杂交过程可在普通96孔板上进行，可方便地使用多通道排枪进行移液操作，整个杂交过程便捷快速，可有效实现高通量基因分型。

主权项

1、一组用于人白细胞抗原基因分型的探针，其特征在于，探针序列如下：第一组HLA-APA01 TGAGGTATTTCTTCACATCCGCATCGCCGTGGGCPA02 GAGGTATTTCTACACCTCTTCATCGCCGTGGGCPA03 CGGTTCGACAGCTGGAGGGGCCGGAGTTACCGAGTGGACCTPA04 GCAGGAGAGGCCTGAGTATTGPA05 AGGAGGGGCCGGAGTAAGGCCCACTCACAGPA06 TATTGGGACCTGCAGACACGGPA07 GACACGGAATATGAAGGCCCAPA08 GGAATGTGAAGGCCCAGTCACPA09 GGCCCACTCACAGACTGACCGPA10 TCACAGATTGACCGAGTGGACPA11 ACAGACTGACCGAGTGGACCTPA12 GAGAGCCTGCGGATCGCGCTPA13 ACCGAGCGAACCTGGGGACCPA14 CGAGCCAGAGGATGGGGCCCAGTCACAGACPA15 TCACAGACTCACCGAGTGGACPA16 GACGAGGAGACAGGGAAAGTGPA17 AGGTATTTCTACACCTCCGTGATCGCAGTGGGCTACPA18 CGGAACACACGGAATCGGATCGCGCTCCGCPA19 TGGGACCAGGAGACATTTTTGGCCCAGTCACAGACPA20 TTGGGACCGGAACACACGGAPA21 AAGGCCCAGTCACAGCGAGTGGACCTGGGPA22 CCTGCGGATCGCGCTCCGCTAPA23 ACACGGAAAGTGAAGGCTTTCGAGTGGACCTGGGPA24 TGGGACCAGGAGACATTTTTCCTGGGGACCCTGCGPA25 GCGGTTCGACAGCCGCGCTCCGCTACTAPA26 TCATCGCAGTGGGCTGTTCGACAGCGACGCPA27 CTACACCTCCGTGTCGGCCCACTCACAGACPA28 GGACCGGAACACACGCGAGCGAACCTGGPA29 CTGACCGAGAGAACCTGGGGAPA30 AGCAGGAGGGTCCGGAGTATTPA31 TTTCTACACCTCCGTGTCTTTAGGCCCAGTCACAGAPA32 GCCCACTCACAGACTCACCGPA33 GGAGACACGGAAAGTGAAGGCPA34 CAGCTCAGACCACCAAGCPA35 GAGGCGGCCCGTGTGGCGGAPA36 CCTACCTGGATGGCACGTGCGPA37 CTGGAGGGCCGGTGCGTGGAPA38 CTGGAGGGCACGTGCGTGGAPA39 CCTGCGCTCTTGGTAGCAGCAGAGAGCTGGAGTGGCTCCPA40 GCGGAGCAGCAGAGAGCCTACPA41 CAAGTGGGAGACGGCCCATGPA42 GGGTCGGACTGGCGCTTCCTPA43 TCGCCCTGAAAGAGGACCTGCPA44 GAGCAGTTGAGAGCCCGTGGAGTGGCTCPA45 GAGCAGTTGAGAGCCTACCTPA46 ACATGGCGGCTCAGAAAGTGGACGGGCTCCGCPA47 GAGCAGTGGAGAGCCTACCTPA48 CGTGGGGTCGGACTTTGGTACCAGCAGGACGCPA49 TACCAGCAGGACGCTTACGACPA50 GTGCGTGGACGGGCTCCGCAGPA51 TACCACCAGTACGCCAGTGGGAGGCGGCCCPA52 GGAGCAGTTGAGAGCGCGTGGACGGGCTCCPA53 TCAGATCACCCAGCGCAAGTGPA54 GGTACCGGCAGGACGAGCACAAGTGGGAGGPA55 TCTCACACCATCCAGAGAGCAGTGGAGAGCCPA56 GGGTACCGGCAGGACGCCTAPA57 CATCGCCCTGAAAGAGGGCGGCTCAGATCACPA58 CGGCCCATGAGGCGTGGAGGGCACGTGCPA59 ACCTGCGCTCTTGGAGACATGGCAGCTGTGGAGTGGCTCPA60 TCTCACACCATCCAGATGATGTATPA61 ACCACCAGTACGCCTACTTTGATCACCAAGCGCAAPA62 CCCATGTGGCGGAGTTTTTCGTGGAGTGGCTCCGAPC2 GCTACTACAACCAGAGCGAGGAPC3 GACGGCAAGGATTACATCGCCANC GACGGCAAGCATTACATCG第二组HLA-BPB01 TGCAAGGCCAAGGCACAGACTPB02 CTGCGCACCGCGCTCCGCTACTACPB03 ACTGACCGAGAGAGCCTGCGGPB04 CTGCGGATCGCGCTCCGCTACPB05 CAGATCTCCAAGACCAACACAPB06 ACACAGATCTACAAGGCCCAGGCAPB07 ACACAGATCTTCAAGACCAACPB08 ACACAGATCTGCAAGACCAACPB09 GCCACGAGTCCGAGGAAGGAGPB10 AGACTTACCGAGAGAGCCTGCGGPB11 CGGACCCTGCTCCGCTACTACPB12 GCCGCGAGTCCGAGAGAGGAGPB13 CGGAACACACAGATCTCCAAGACCPB14 TGGGACCGGGAGACACAGATCPB15 ATGAAGGCCTCCGCGCAGACTPB16 CGGGAGACACAGATCTCCAAGACCPB17 CCGAGGACGGAGCCCCGGGCGPB18 GAGGTATTTCCACACCGCCATPB19 ACACCGCCATGTCCCTCCTGCGGAACCTGCGPB20 CGTGGATAGAGCAGGTTTGGCCCAGGCACAGACPB21 AGACCAACACACAGACTGACCPB22 ACGACACCCAGTTCGATTCGAGAGAGGAGCCGCPB23 CTTCATCGCAGTGGGTTACCGAGAGAGGAGCCGPB24 GACGACACCCAGTTCGATTCCGGGAGACACAGATPB25 ACGACACCCAGTTCGATACCGAGGATGGCGCCCPB26 TGCGGAACCTGCGCGGCTACPB27 ACGACACGCAGTTCGAATGCGGATCGCGCTCCPB28 ACACAGATCTGCAAGCGGATCGCGCTCCGCPB29 GCTTCATTGCAGTGGTTTTTCGGATCGCGCTCCGCPB30 CGGAACACACAGATCCGGATCGCGCTCCGCPB31 CGGGAGACACAGATCCGGATCGCGCTCCGCPB32 GTATTTCCACACCGCCAAGCCTGCGGAACCTGPB33 ACGACACCCAGTTCGCAAGGCCCAGGCACAPB34 GCCGTGGGTGGAGCAGGAGPB35 AGAAGTACAAGCGCCAGGCACPB36 CAGATCTCCAAGACCAACATCGCGCTCCGCPB37 GACGCCACGAGTCCGATGGCGCCCCGPB38 CCGAGGATGGCGCCCCCGGAACACACAGATPB39 AGTCCGAGGATGGCGCCCCTGCGGATCGCGCTCPB40 GACTTACCGAGAGAACCTGCPB41 ATTTCTACACCTCCGTGTTGCGGAACCTGCGCGPB42 GTCCGAGAGAGGAGCTTTGACCGGGAGACACAGPB43 GCTTCATCTCAGTGGTTTCAAGGCCCAGGCACAPB44 ACACCTCCGTGTCCTCATCACCGTGGGACTACAACCAGPB45 ACCGGGAGACACAGAATTCCTGCGGAACCTGCGPB46 GTCCGAGGAAGGAGCTTATGCGGAACCTGCGCGPB47 CCGAGAGAGGAGCTTCCGGAACACACTGGAACCTGCGCGPB48 CCGAGAGAGGAGCTTCGGAACACACTGGAACCTGCGCGPB49 TTCATCGCAGTGGCCGGAACACACATCTACAACCAGPB50 CAGATCTACAAGTTTCCAACACACAGAGCTACAACCAGAGCPB51 GGCGAGTGCGTGGAGTGGCTCPB52 GTGCGTGGACGGGCTCCGCAPB53 CAGAGGATGTCTGGCTGCGACPB54 GAGCAGCTGAGAACCTACCTGPB55 GGGCATGACCAGTACGCCTACPB56 GTGGCGGAGCAGGACAGAGCCTACPB57 CGGAGCAGCGGAGAGCCTACCPB58 ATCTCCCAGCGCAAGTTGGAGPB59 CGCGGGTATGACCAGGACGCCPB60 CGCGGGTACCACCAGGACGCCPB61 CGCGGGCATAACCAGTTCGCCTACPB62 AAGGACACGCTGGAGCGCGCGPB63 CGTGAGGCGGAGCAGTGGAGAPB64 CTGGAGGGCACGTGCGTGGAGPB65 CGGCTGCGACCTGGGGCCGGAPB66 GGGCATGACCAGTCCGCCTACGACPB67 GGGTCTCACACCCTCCAGAGGATGPB68 CGTGAGGCGGAGCAGCTGAGAPB69 TGCGTGGAGTCGCTCCGCAGAPB70 CGCGGCGGACACGGCGGCTCPB71 CAGAGGATGTACGGCTGCGACPB72 ACCAGTTAGCCTACGGGACCTGAGCTCCTGPB73 GGACCTGAGCTCCTGTTTGTGGAGTGGCTCCGCPB74 CGGACACGGCGGCTCTGGGAAGGAGACGCTGPB75 CTCCAGAGCATGTACGGGCATAACCAGTACGCCPB76 GCGGGCATGACCAGTGACGGACACCGCGGCTCPB77 CATGACCAGTCCGCCTTTTCGCAGATACCTGGAGPB78 GGGCATAACCAGTACGCCTACPB79 CGGAGCAGGACAGAGATAGGAGTCGCTCCGCAGPB80 GATGTATGGCTGCGACTGACCAGTCCGCCTAPB81 CACACTTGGCAGACGTTAGGAGGGCCTGTGCGTPB82 CATCGCCCTGAACGTTTGACCTGCGCTCCTGPB83 CACCCTCCAGAGGATGTGAGGGCACGTGCGTGPB84 AGCAGCTGAGAGCCTTTTCGCAGACACCTGGAGPB85 GAGCAGCTGAGAGCCTACCTGPB86 GAGGGCCTGTGCGTGTTTCGCAGACACCTGGAGPB87 CCTCCAGAGCATGTACGTTTGCTCAGATCTCCCAGCGPB88 GGTCTCACACTTGGCAGAATTCTGCGACCTGGGGCCPB89 TCCAGGTGATGTATGGCTGCGPB90 CATGACCAGTTCGCCTACGACPB91 GCGGACACGGCGGCTGAGCAGCGGAGAGCCPB92 ACATCATCCAGAGGATGACCCTGAACGAGGACCPB93 GTGAGGCGGAGCAGCGGAGAPB94 CTGCGACCTGGGGCTACATCGCCCTGAATCCCGTGTGGCGGAGPB95 CAGCTGAGAGCCTACAAAAGGGCCTGTGCGTGGPB96 TCCAGAGCATGTACGGCTGCGBPC2 ACTACAACCAGAGCGAGGCCGBPC3 ATTACATCGCCCTGAACGAGGACBNC CATCGCCGTCAACGAGGAC第三组HLA-DRB1PD01 AGCTTAAGTTTGAATGTCATTTCTPD02 GAGCAGGTTAAACATGAGTGTCATPD03 GTGGCAGGGTAAGTATAAGTGPD04 GAAGCAGGATAAGTTTGAGTGPD05 GGAGGAGGTTAAGTTTGAGTGPD06 ATAACCAGGAGGAGTCCGTGCPD07 CAGAAGGACATCCTGGAAGACPD08 GCGGTGACGGAGCTTTTCTGCTGCGGAGCACTPD09 GAGTACTCTACGTCTGTTTTGGAGCAGAAGCGGGPD10 GGGCCCTGGTGGACTTTGTTGTG GAGAGCTTCPD11 GGACATCCTGGAAGACGAGCPD12 AGTACTCTACGTCTGATTTCGGCCTGATGAGGAGPD13 GGAGTACGTGCGCTTTTTGGCCCTGGTGGACAPD14 GTGCGGTTACTGGAGAGACACPD15 GCCTGATGCTGAGTACTGGAPD16 CCTGATGAGGAGTACTTTTCTGGAAGACGAGCGGPD17 GACATCCTGGAAGACAAGCGGPD18 GGGAGTTCCGGGCTTACCTGGAAGACGAGCGPD19 GAGGAGTTCGTGCGCATTCCTGATGAGGAGTACPD20 TGATGAGGAGCACTGGAACAGPD21 AGATACTTCTATCACCAAGAGGAGPD22 GAGGAGAACGTGCGCATTGTGGACAACTACTGCPD23 ACTCTACGGGTGAGTGTATTGGGAGTACCGGGCPD24 GTTCCTGGACAGATACTATTCTGATGCCGAGTACTPD25 CAGAAGGACCTCCTGGAAGACPD26 GGAGTACTCTACGTCTGTTTGTGGACAATTACTGCAPD27 TGGAGTACTCTACGTCTTTTGCCTG ACGCTGAGTAPD28 GGGTGAGTGTTATTTCTTCAATGGPD29 CCTGGACAGATACCTTCGACAGCGACCCTGATGCGGAGPD30 CTACGTCTGAGTGCCTTCGACAGCGACTGGGTTGTGGAGAGPD31 CTACGTCTGAGTGGGAGTTCCGGGCAAGGACCTCCTGGPD32 GTACTCTACGTCTGAGTGCGGTACCTGGACPD33 GACATCCTGGAGGACAGGCGPD34 CTTGGAGTACTCTACGCTTCCATAACCAGGAGPD35 CCTGGAGAGATACTTCGAGGAGAACGTGCGCPD36 AGGAGAACGTGCGCTCCTGATGCCGAGTACTGPD37 GAGGAGTTCGTGCGCTGCTGCGGAGCACTPD38 GAAGACGAGCGGGCGGTTGGTGAGAGCTTCPD39 GGTTCCTGGACAGATACGCCTAGCGCCGAGTPD40 GGAGTACTCTACGGGTGAGTGPD41 TCTACGTCTGAGTGTCCCTGGAAGACAGGCGPD42 CCTGGAAGACAGGCGACGGGGTTGTGGAGAPD43 CTGATGCCGAGTACTGCCTGGAAGACAGGCPD44 TGGCAGCCTAAGAGGTCCTGGAAGACAGGCGPD45 GAGGAGTCCGTGCGCGGGAGTTCCGGGCPD46 GGCCTAGCGCCGATGGAGCAGAGGCGGGPD47 GGTTCCTGGACAGATACGAAGGACTTCCTGGAPD48 GGAGTACGTGCGCTTCCTGATGAGGAGTACTGPD49 GGAGAGATACTTCCATAGTGGACACCTACTGCPD50 CAGGAGGAGAACGTGCGCCTGGAAGACGAGCGGPD51 TCCTGGAAGACGAGCGGGGGTTGGTGAGAGCPD52 CGGCCTAGCGCCGAGCTGGAGCAGAGGCGGPD53 CTGGAGCAGGCGCGGGGGGTTGGTGAGAGCPD54 TTCTTCAATGGGACGGTTTTTGGACTTCCTGGAAGACPD55 GGTGCGGTTCCTGTGAGTACGTGCGCCGACGTGGGGGAPD56 GCGGTTCCTGGACAGTTTTTGGCCTGATGCCGAGTPD57 GGAGTACGTGCGCATGATGAGGAGTACGAAGGACTTCCTGGAPD58 CTTCGACAGCGAGCCGAGTACTGGACTGGAAGACGAGCPD59 AGGAGTTCGTGCGCCCTGATGCCGAGAGGGTTGTGGAGAPD60 TGGAGTACTCTACGATCGACAGCGACTGGGGCTGTGGAGAGPD61 GTACTCTACGTCTGATTTGTGCGGTACCTGGACPD62 AGGAGAACGTGCGCTATTCCTGATGCCGAGTACTGPD63 TCCTGGACAGATACTATTCTGGAGCAGATTTGTGGACACCTACTGPD64 GAGTACTCTACGTCTGTTTTTCTGGAGCAGAGGCGGPD65 GTACTCTACGTCTGAGTTGATGAGGAGTACTGGPD66 GGCCTGATGCCGAGTTTTTTGGGCCCTGGTGGACAPD67 GTACTCTACGGGTGAGTCGCTTCGACAGCGACPD68 CTCTACGTCTGAGTGTCTTACCTGGAAGACAGGCPD69 CTGATGCCGAGTACTGTTACCTGGAAGACAGGCPD70 CTGGACAGATACTTACTTCGACAGCGATAGGACATCCTGGAPD71 AGAGGAGTACGTGCGTTCCTGGAAGACAGGCPD72 GGTTCCTGGACAGATACATTGAAGGACTTCCTGGAPD73 GAGTACTCTACGTCTGATTTTTCTGGAGCAGAAGCGGPD74 TTCTTCAATGGGACGGGGACTTCCTGGAAGACPD75 CCTGGAGAGATACTTCGGAAGACGAGCGGGCPD76 CCTGGACAGATAAGAGCAGAAGCGGGTCACCTACTGCAGPD77 GCTTCGACAGCGACTGGCCTGTCGCCGATGGAAGACAGGCGPD78 TCTACGTCTGAGTGTCTTTTTCCGCGGTGGACACCTPD79 CTGATGCCGAGTACTCTGGAAGACAGGCGGPD80 CTAAGAGGGAGTGTGGGAGTACCGGGCAAGACAGGCGPD81 GCTTCGACAGCGACTCTGATGCCGAGTGAGCAGAGGCGGGPD82 AGCCTAAGAGGGACTTCGACAGCGACTGAGCAGGCGCGGGCPD83 CCTGGAGAGATACGAGCAGAGGCGGGGACACCTACTGCPD84 CCTGGACAGATACGATGCCGAGTACTAGGTTGGTGAGAGCPD85 TGCGGTTCCTGTGAGGAGTACGTGCGTACGTGGGGGAPD86 GCAGCCTAAGAGGGATTTGGGGAGTACCGGGCGPD87 CTGGACAGATACATTCGACAGCGATCTGGAGCGGAGGCPD88 TCGACAGCGACGTAGGGCCCTGGTGGAGGTTGTGGAGAGCPD89 CTTCGACAGCGTGGGAGTTCCGGGCAAGGACTTCCTGGPD90 TCTACGTCTGAGTTTCGACAGCGACTGAGCAGAAGCGGGDPC GCGCTTCGACAGCGACGTGGGDNC CGCTTCGACTCCGACGTGG