背景：传统的HLA测序分型技术都是对父系母系双倍体进行扩增,由于HLA多样性高,产生了大量的模棱两可难题,而采用基于组特异性单倍体全长测序分型方法来解决模棱两可结果的确认定型鲜有报道。目的：分析基于组特异性单倍体全长测序分型方法对HLA基因分型模棱两可结果准确定型的效果。方法：对2例模棱两可结果标本以低分辨率分型方法结果作为参考起点,通过组特异性扩增分离,基于等位基因全长单倍体的Sanger测序(PCR-SBT)分型。

王宋兴，等. 基于HLA 组特异性单倍体全长测序方法对模棱两可结果的准确定型 P .O. Box 10002, Shenyang 110180

3212

www.

CRTER

.org

Exon1 18 A*11:01:01:01 CGGTCGCTCT TCTAAAGTCC GCACGCACCC ACCGGGACTC AGATTCTCCC CAGACCCCGA GGATGGCCGT CATGGCGCCC A*11:01:01:01NEW ---------- ---------- ---------- ---------- ---------- ---------- ---------- ---------- Intron1 98 A*11:01:01:01 CGAACCCTCC TCCTGCTACT CTCGGGGGCC CTGGCCCTGA CCCAGACCTG GGCGGGTGAG TGCGGGGTCG GGAGGGAAAC A*11:01:01:01NEW ---------- ---------- ---------- ---------- ---------- ---------- ---------- ---------- 178 A*11:01:01:01 CGCCTCTGCG GGGAGAAGCA AGGGGCCCTC CTGGCGGGGG CGCAGGACCG GGGGAGCCGC GCCGGGAGGA GGGTCGGGCA A*11:01:01:01NEW ---------- ---------- ---------- ---------- ---------- ---------- ---------- ---------- Exon2 258 A*11:01:01:01 GGTCTCAGCC ACTGCTCGCC CCCAGGCTCC CACTCCATGA GGTATTTCTA CACCTCCGTG TCCCGGCCCG GCCGCGGGGA A*11:01:01:01NEW ---------- ---------- ---------- ---------- ---------- ---------- ---------- ---------- 338 A*11:01:01:01 GCCCCGCTTC ATCGCCGTGG GCTACGTGGA CGACACGCAG TTCGTGCGGT TCGACAGCGA CGCCGCGAGC CAGAGGATGG A*11:01:01:01NEW ---------- ---------- ---------- ---------- ---------- ---------- ---------- ---------- 418 A*11:01:01:01 AGCCGCGGGC GCCGTGGATA GAGCAGGAGG GGCCGGAGTA TTGGGACCAG GAGACACGGA ATGTGAAGGC CCAGTCACAG A*11:01:01:01NEW ---------- ---------- ---------- ---------- ---------- ---------- ---------- ---------- Intron2 498 A*11:01:01:01 ACTGACCGAG TGGACCTGGG GACCCTGCGC GGCTACTACA ACCAGAGCGA GGACGGTGAG TGACCCCGGC CCGGGGCGCA A*11:01:01:01NEW ---------- ---------- ---------- ---------- ---------- ---------- ---------- ---------- 578 A*11:01:01:01 GGTCACGACC CCTCATCCCC CACGGACGGG CCAGGTGGCC CACAGTCTCC GGGTCCGAGA TCCACCCCGA AGCCGCGGGA A*11:01:01:01NEW ---------- ---------- ---------- ---------- ---------- ---------- ---------- ---------- 658 A*11:01:01:01 CCCCGAGACC CTTGCCCCGG GAGAGGCCCA GGCGCCTTTA CCCGGTTTCA TTTTCAGTTT AGGCCAAAAA TCCCCCCGGG A*11:01:01:01NEW ---------- ---------- ---------- ---------- ---------- ---------- ---------- ---------- Exon3 738 A*11:01:01:01 TTGGTCGGGG CCGGGCAGGG CTTGGGGGAC TGGGCTGACC GCGGGGTCGG GGCCAGGTTC TCACACCATC CAGATAATGT A*11:01:01:01NEW ---------- ---------- ---------- ---------- ---------- ---------- ---------- ---------- 818 A*11:01:01:01 ATGGCTGCGA CGTGGGGCCG GACGGGCGCT TCCTCCGCGG GTACCGGCAG GACGCCTACG ACGGCAAGGA TTACATCGCC A*11:01:01:01NEW ---------- ---------- ---------- ---------- ---------- ---------- ---------- --------T- 898 A*11:01:01:01 CTGAACGAGG ACCTGCGCTC TTGGACCGCG GCGGACATGG CAGCTCAGAT CACCAAGCGC AAGTGGGAGG CGGCCCATGC A*11:01:01:01NEW ---------- ---------- ---------- ---------- ---------- ---------- ---------- ---------- 978 A*11:01:01:01 GGCGGAGCAG CAGAGAGCCT ACCTGGAGGG CCGGTGCGTG GAGTGGCTCC GCAGATACCT GGAGAACGGG AAGGAGACGC A*11:01:01:01NEW ---------- ---------- ---------- ---------- ---------- ---------- ---------- ---------- Intron3 1058 A*11:01:01:01 TGCAGCGCAC GGGTACCAGG GGCCACGGGG CGCCTCCCTG ATCGCCTATA GATCTCCCGG GCTGGCCTCC CACAAGGAGG A*11:01:01:01NEW ---------- ---------- ---------- ---------- ---------- ---------- ---------- ---------- 1138 A*11:01:01:01 GGAGACAATT GGGACCAACA CTAGAATATC ACCCTCCCTC TGGTCCTGAG GGAGAGGAAT CCTCCTGGGT TTCCAGATCC A*11:01:01:01NEW ---------- ---------- ---------- ---------- ---------- ---------- ---------- ---------- 1218 A*11:01:01:01 TGTACCAGAG AGTGACTCTG AGGTTCCGCC CTGCTCTCTG ACACAATTAA GGGATAAAAT CTCTGAAGGA GTGACGGGAA A*11:01:01:01NEW ---------- ---------- ---------- ---------- ---------- ---------- ---------- ---------- 1298 A*11:01:01:01 GACGATCCCT CGAATACTGA TGAGTGGTTC CCTTTGACAC CGGCAGCAGC CTTGGGCCCG TGACTTTTCC TCTCAGGCCT A*11:01:01:01NEW ---------- ---------- ---------- ---------- ---------- ---------- ---------- ---------- 1378 A*11:01:01:01 TGTTCTCTGC TTCACACTCA ATGTGTGTGG GGGTCTGAGT CCAGCACTTC TGAGTCTCTC AGCCTCCACT CAGGTCAGGA A*11:01:01:01NEW ---------- ---------- ---------- ---------- ---------- ---------- ---------- ---------- 1458 A*11:01:01:01 CCAGAAGTCG CTGTTCCCTT CTCAGGGAAT AGAAGATTAT CCCAGGTGCC TGTGTCCAGG CTGGTGTCTG GGTTCTGTGC A*11:01:01:01NEW ---------- ---------- ---------- ---------- ---------- ---------- ---------- ---------- 1538 A*11:01:01:01 TCTCTTCCCC ATCCCGGGTG TCCTGTCCAT TCTCAAGATG GCCACATGCG TGCTGGTGGA GTGTCCCATG ACAGATGCAA A*11:01:01:01NEW ---------- ---------- ---------- ---------- ---------- ---------- ---------- ---------- Exon4 1618 A*11:01:01:01 AATGCCTGAA TTTTCTGACT CTTCCCGTCA GACCCCCCCA AGACACATAT GACCCACCAC CCCATCTCTG ACCATGAGGC A*11:01:01:01NEW ---------- ---------- ---------- ---------- ---------- ---------- ---------- ---------- 1698 A*11:01:01:01 CACCCTGAGG TGCTGGGCCC TGGGCTTCTA CCCTGCGGAG ATCACACTGA CCTGGCAGCG GGATGGGGAG GACCAGACCC A*11:01:01:01NEW ---------- ---------- ---------- ---------- ---------- ---------- ---------- ---------- 1778 A*11:01:01:01 AGGACACGGA GCTCGTGGAG ACCAGGCCTG CAGGGGATGG AACCTTCCAG AAGTGGGCGG CTGTGGTGGT GCCTTCTGGA A*11:01:01:01NEW ---------- ---------- ---------- ---------- ---------- ---------- ---------- ---------- Intron4 1858 A*11:01:01:01 GAGGAGCAGA GATACACCTG CCATGTGCAG CATGAGGGTC TGCCCAAGCC CCTCACCCTG AGATGGGGTA AGGAGGGAGA A*11:01:01:01NEW ---------- ---------- ---------- ---------- ---------- ---------- ---------- ---------- 1938 A*11:01:01:01 TGGGGGTGTC ATGTCTCTTA GGGAAAGCAG GAGCCTCTCT GGAGACCTTT AGCAGGGTCA GGGCCCCTCA CCTTCCCCTC A*11:01:01:01NEW ---------- ---------- ---------- ---------- ---------- ---------- ---------- ---------- Exon5 2018 A*11:01:01:01 TTTTCCCAGA GCTGTCTTCC CAGCCCACCA TCCCCATCGT GGGCATCATT GCTGGCCTGG TTCTCCTTGG AGCTGTGATC A*11:01:01:01NEW ---------- ---------- ---------- ---------- ---------- ---------- ---------- ---------- Intron5 2098 A*11:01:01:01 ACTGGAGCTG TGGTCGCTGC CGTGATGTGG AGGAGGAAGA GCTCAGGTGG AGAAGGGGTG AAAGGTGGGG TCTGAGATTT A*11:01:01:01NEW ---------- ---------- ---------- ---------- ---------- ---------- ---------- ---------- 2178 A*11:01:01:01 CTTGTCTCAC TGAGGGTTCC AAGCCCCAGC TAGAAATGTG CCCTGTCTCA TTACTGGGAA GCACCTTCCA CAATCATGGG A*11:01:01:01NEW ---------- ---------- ---------- ---------- ---------- ---------- ---------- ---------- 2258 A*11:01:01:01 CCGACCCAGC CTGGGCCCTG TGTGCCAGCA CTTACTCTTT TGTAAAGCAC CTGTTAAAAT GAAGGACAGA TTTATCACCT A*11:01:01:01NEW ---------- ---------- ---------- ---------- ---------- ---------- ---------- ---------- 2338 A*11:01:01:01 TGATTACGGC GGTGATGGGA CCTGATACCA GCAGTCACAA GTCACAGGGG AAGGTCCCTG AGGACAGACC TCAGGAGGGC A*11:01:01:01NEW ---------- ---------- ---------- ---------- ---------- ---------- ---------- ---------- 2418 A*11:01:01:01 TATTGGTCCA GGACCCACAC CTGCTTTCTT CATGTTTCCT GATCCCGCCC TGGGTCTGCA GTCACACATT TCTGGAAACT A*11:01:01:01NEW ---------- ---------- ---------- ---------- ---------- ---------- ---------- ---------- 2498 A*11:01:01:01 TCTCTGGGGT CCAAGACTAG GAGGTTCCTC TAGGACCTTA AGGCCCTGGC TCCTTTCTGG TATCTCACAG GACATTTTCT A*11:01:01:01NEW ---------- ---------- ---------- ---------- ---------- ---------- ---------- ---------- Exon6 Intron6 2578 A*11:01:01:01 TCCCACAGAT AGAAAAGGAG GGAGTTACAC TCAGGCTGCA AGTAAGTATG AAGGAGGCTG ATGCCTGAGG TCCTTGGGAT A*11:01:01:01NEW ---------- ---------- ---------- ---------- ---------- ---------- ---------- ----------

2658 A*11:01:01:01 ATTGTGTTTG GGAGCCCATG GGGGAGCTCA CCCACCCCAC AATTCCTCCT CTAGCCACAT CTTCTGTGGG ATCTGACCAG A*11:01:01:01NEW ---------- ---------- ---------- ---------- ---------- ---------- ---------- ---------- Exon7 Intron7 2738 A*11:01:01:01 GTTCTGTTTT TGTTCTACCC CAGGCAGTGA CAGTGCCCAG GGCTCTGATG TGTCTCTCAC AGCTTGTAAA GGTGAGAGCT A*11:01:01:01NEW ---------- ---------- ---------- ---------- ---------- ---------- ---------- ---------- 2818 A*11:01:01:01 TGGAGGGCCT GATGTGTGTT GGGTGTTGGG CGGAACAGTG GACACAGCTG TGCTATGGGG TTTCTTTGCA TTGGATGTAT A*11:01:01:01NEW ---------- ---------- ---------- ---------- ---------- ---------- ---------- ---------- 2898 A*11:01:01:01 TGAGCATGCG ATGGGCTGTT TAAGGTGTGA CCCCTCACTG TGATGGATAT GAATTTGTTC ATGAATATTT TTTTCTATAG A*11:01:01:01NEW ---------- ---------- ---------- ---------- ---------- ---------- ---------- ---------- Exon8 2978 A*11:01:01:01 TGTGAGACAG CTGCCTTGTG TGGGACTGAG AGGCAAGAGT TGTTCCTGCC CTTCCCTTTG TGACTTGAAG AACCCTGACT A*11:01:01:01NEW ---------- ---------- ---------- ---------- ---------- ---------- ---------- ---------- 2985 A*11:01:01:01 TTGTTTC

A*11:01:01:01NEW -------

-234 C*08:01:01 TGACAAAGAT GCTTGGTGTA GGAGAAGAGG GATCAGGACG AAGTCCCAGG TCCCGGGCGG GGCTCTCAGG GTCTCAGGCT C*08:01:01NEW ---------- ---------- ---------- ---------- ---------- ---------- ---------- ---------- -154 C*08:01:01 CCAAGGGCCG TGTCTGCACT GGGGAGGCGC CGCGTTGAGG ATTCTCCACT CCCCTGAGTT TCACTTCTTC TTCCAACCTG C*08:01:01NEW ---------- ---------- ---------- ---------- ---------- ---------- ---------- ---------- -74 C*08:01:01 CGTCGGGTCC TTCTTCCTGA ATACTCATGA CGCGTCCCCA ATTCCCACTC CCATTGGGTG TCGGGTTCTA GAGAAGCCAA C*08:01:01NEW ---------- ---------- ---------- ---------- ---------- ---------- ---------- ---------- Exon1 7 C*08:01:01 TCAGCGTCTC CGCAGTCCCG GTTCTAAAGT CCCCAGTCAC CCACCCGGAC TCGGATTCTC CCCAGACGCC GAGATGCGGG C*08:01:01NEW ---------- ---------- ---------- ---------- ---------- ---------- ---------- ---------- Intron1 87 C*08:01:01 TCATGGCGCC CCGAACCCTC ATCCTGCTGC TCTCGGGAGC CCTGGCCCTG ACCGAGACCT GGGCCTGTGA GTGCGAGGTT C*08:01:01NEW ---------- ---------- ---------- ---------- ---------- ---------- ---------- ---------- 167 C*08:01:01 GGGAGGGAAA CGGCCTCTGC GGAGAGGAGC GAGGGGCCCG CCCGGCGAGG GCGCAGGACC CGGGGAGCCG CGCAGGGAGG C*08:01:01NEW ---------- ---------- ---------- ---------- ---------- ---------- ---------- ---------- Exon2 247 C*08:01:01 AGGGTCGGGC GGGTCTCAGC CCCTCCTCGC CCCCAGGCTC CCACTCCATG AGGTATTTCT ACACCGCCGT GTCCCGGCCC C*08:01:01NEW ---------- ---------- ---------- ---------- ---------- ---------- ---------- ---------- 327 C*08:01:01 GGCCGCGGAG AGCCCCGCTT CATCGCAGTG GGCTACGTGG ACGACACGCA GTTCGTGCAG TTCGACAGCG ACGCCGCGAG C*08:01:01NEW ---------- ---------- ---------- ---------- ---------- ---------- ---------- ---------- 407 C*08:01:01 TCCAAGAGGG GAGCCGCGGG CGCCGTGGGT GGAGCAGGAG GGGCCGGAGT ATTGGGACCG GGAGACACAG AAGTACAAGC C*08:01:01NEW ---------- ---------- ---------- ---------- ---------- ---------- ---------- ---------- Intron2 487 C*08:01:01 GCCAGGCACA GACTGACCGA GTGAGCCTGC GGAACCTGCG CGGCTACTAC AACCAGAGCG AGGCCGGTGA GTGACCCCGG C*08:01:01NEW ---------- ---------- ---------- ---------- ---------- ---------- ---------- ---------- 567 C*08:01:01 CCCGGGGCGC AGGTCCCGAC CCCTCCCCAT CCCCCACGGA CGGCCCGGGT CGCCCCGAGT CTCCCGGTCT GAGATCCACC C*08:01:01NEW ---------- ---------- ---------- ---------- ---------- ---------- ---------- ---------- 647 C*08:01:01 CCGAGGCTGC GGAACCCGCC CAGACCCTCG ACCGGAGAGA GCCCCAGTCA CCTTTACCCG GTTTCATTTT CAGTTTAGGC C*08:01:01NEW ---------- ---------- ---------- ---------- ---------- ---------- ---------- ---------- Exon3 727 C*08:01:01 CAAAATCCCC GCGGGTTGGT CGGGGCTGGG GCGGGGCTCG GGGGACGGGG CTGACCACGG GGGCGGGGCC AGGGTCTCAC C*08:01:01NEW ---------- ---------- ---------- ---------- ---------- ---------- ---------- ---------- 807 C*08:01:01 ACCCTCCAGA GGATGTATGG CTGCGACCTG GGGCCCGACG GGCGCCTCCT CCGCGGGTAT AACCAGTTCG CCTACGACGG C*08:01:01NEW ---------- ---------- ---------- ---------- ---------- ---------- ---------- ---------- 887 C*08:01:01 CAAGGATTAC ATCGCCCTGA ATGAGGACCT GCGCTCCTGG ACCGCCGCGG ACACGGCGGC TCAGATCACC CAGCGCAAGT C*08:01:01NEW ---------- ---------- ---------- ---------- ---------- ---------- ---------- -G-------- 967 C*08:01:01 GGGAGGCGGC CCGTACGGCG GAGCAGCTGA GAGCCTACCT GGAGGGCACG TGCGTGGAGT GGCTCCGCAG ATACCTGGAG C*08:01:01NEW ---------- ---------- ---------- ---------- ---------- ---------- ---------- ---------- Intron3 1047 C*08:01:01 AACGGGAAGA AGACGCTGCA GCGCGCGGGT ACCAGGGGCA GTGGGGAGCC TTCCCCATCT CCTGTAGATC TCCCGGGATG C*08:01:01NEW ---------- ---------- ---------- ---------- ---------- ---------- ---------- ---------- 1127 C*08:01:01 GCCTCCCACG AGGAGGGGAG GAAAATGGGA TCAGCGCTGG AATATCGCCC TCCCTTGAAT GGAGAATGGG ATGAGTTTTC C*08:01:01NEW ---------- ---------- ---------- ---------- ---------- ---------- ---------- ---------- 1207 C*08:01:01 CTGAGTTTCC TCTGAGGGCC CCCTCTGCTC TCTAGGACAA TTAAGGGATG AAGTCCTTGA GGAAATGGAG GGGAAGACAG C*08:01:01NEW ---------- ---------- ---------- ---------- ---------- ---------- ---------- ---------- 1287 C*08:01:01 TCCCTGGAAT ACTGATCAGG GGTCCCCTTT GACCACTTTG ACCACTGCAG CAGCTGTGGT CAGGCTGCTG ACCTTTCTCT C*08:01:01NEW ---------- ---------- ---------- ---------- ---------- ---------- ---------- ---------- 1367 C*08:01:01 CAGGCCTTGT TCTCTGCCTC ACGCTCAATG TGTTTAAAGG TTTGATTCCA GCTTTTCTGA GTCCTTCGGC CTCCACTCAG C*08:01:01NEW ---------- ---------- ---------- ---------- ---------- ---------- ---------- ---------- 1447 C*08:01:01 GTCAGGACCA GAAGTCGCTG TTCCTCCCTC AGAGACTAGA ACTTTCCAAT GAATAGGAGA TTATCCCAGG TGCCTGTGTC C*08:01:01NEW ---------- ---------- ---------- ---------- ---------- ---------- ---------- ---------- 1527 C*08:01:01 CAGGCTGGCG TCTGGGTTCT GTGCCCCCTT CCCCACCCCA GGTGTCCTGT CCATTCTCAG GATGGTCACA TGGGCACTGT C*08:01:01NEW ---------- ---------- ---------- ---------- ---------- ---------- ---------- ---------- Exon4 1607 C*08:01:01 TGGAGTGTCG CAAGAGAGAT ACAAAGTGTC TGAATTTTCT GACTCTTCCC GTCAGAACAC CCAAAGACAC ACGTGACCCA C*08:01:01NEW ---------- ---------- ---------- ---------- ---------- ---------- ---------- ---------- 1687 C*08:01:01 CCATCCCGTC TCTGACCATG AGGCCACCCT GAGGTGCTGG GCCCTGGGCT TCTACCCTGC GGAGATCACA CTGACCTGGC C*08:01:01NEW ---------- ---------- ---------- ---------- ---------- ---------- ---------- ---------- 1767 C*08:01:01 AGCGGGATGG CGAGGACCAA ACTCAGGACA CCGAGCTTGT GGAGACCAGG CCAGCAGGAG ATGGAACCTT CCAGAAGTGG C*08:01:01NEW ---------- ---------- ---------- ---------- ---------- ---------- ---------- ---------- 1847 C*08:01:01 GCAGCTGTGG TGGTGCCTTC TGGAGAaGAG CAGAGATACA CGTGCCATGT GCAGCACGAG GGGCTGCCAG AGCCCCTCAC C*08:01:01NEW ---------- ---------- ---------- ---------- ---------- ---------- ---------- ---------- Intron4 1927 C*08:01:01 CCTGAGATGG GGTAAGGAGG GGGATGAGGG GTCATGTGTC TTCTCAGGGA AAGCAGAAGT CCTGGAGCCC TTCAGCCGGG C*08:01:01NEW ---------- ---------- ---------- ---------- ---------- ---------- ---------- ---------- Exon5 2027 C*08:01:01 TCAGGGCTGA GGCTTGGGGG TCAGGGCCCC TCACCTTCCC CTCCTTTCCC AGGGCCATCT TCCCAGCCCA CCATCCCCAT C*08:01:01NEW ---------- ---------- ---------- ---------- ---------- ---------- ---------- ---------- 2107 C*08:01:01 CGTGGGCATC GTTGCTGGCC TGGCTGTCCT GGCTGTCCTA GCTGTCCTAG GAGCTGTGAT GGCTGTTGTG ATGTGTAGGA C*08:01:01NEW ---------- ---------- ---------- ---------- ---------- ---------- ---------- ---------- Intron5 2187 C*08:01:01 GGAAGAGCTC AGGTAGGGAA GGGGTGAGGA GTGGGGTCTG GGTTTTCTTG TCCCACTGGG AGTTTCAAGC CCCAGGTAGA C*08:01:01NEW ---------- ---------- ---------- ---------- ---------- ---------- ---------- ---------- 2247 C*08:01:01 AGTGTGCCCC ACCTCGTTAC TGGAAGCACC ATCCACACAT GGGCCATCCC AGCCTGGGAC CCTGTGTGCT AGCACTTACT C*08:01:01NEW ---------- ---------- ---------- ---------- ---------- ---------- ---------- ---------- 2327 C*08:01:01 CTGTTGTGAA GCACATGACA ATGAAGGACA GATGTATCAC CTTGATGATT ATGGTGTTGG GGTCCTTGAT TCCAGCATTC C*08:01:01NEW ---------- ---------- ---------- ---------- ---------- ---------- ---------- ----------

2407

C*08:01:01 ATGAGTCAGG GGAAGGTCCC TGCTAAGGAC AGACCTTAGG AGGGCAGTTG CTCCAGAACC CACAGCTGCT TTCCCCGTGT C*08:01:01NEW ---------- ---------- ---------- ---------- ---------- ---------- ---------- ---------- 2487 C*08:01:01 TTCCTGATCC TGCCCTGGGT CTGCAGTCAT AGTTCTGGAA ACTTCTCTTG GGTCCAAGAC TAGGAGGTTC CCCTAAGATC C*08:01:01NEW ---------- ---------- ---------- ---------- ---------- ---------- ---------- ---------- Exon6 2567 C*08:01:01 GCATGGCCCT GCCTCCTCCC TGTCCCCTCA CAGGGCATTT TCTTCCCACA GGTGGAAAAG GAGGGAGCTG CTCTCAGGCT C*08:01:01NEW ---------- ---------- ---------- ---------- ---------- ---------- ---------- ---------- Intron6 2647 C*08:01:01 GCGTGTAAGT GATGGCGGTG GGCGTGTGGA GGAGCTCACC CACCCCATAA TTCCTCTTGT CCCACATCTC CTGCGGGCTC C*08:01:01NEW ---------- ---------- ---------- ---------- ---------- ---------- ---------- ---------- Exon7 Intron7 C*08:01:01 TGACCAGGTC TTTTTTTTTG TTCTACCCCA GCCAGCAACA GTGCCCAGGG CTCTGATGAG TCTCTCATCG CTTGTAAAGG C*08:01:01NEW ---------- ---------- ---------- ---------- ---------- ---------- ---------- ---------- 2807 C*08:01:01 TGAGATTCTG GGGAGCTGAA GTGGTCGGGG GTGGGGCAGA GGGAAAAGGC CTAGGTAATG GGGATCCTTT GATTGGGACG C*08:01:01NEW ---------- ---------- ---------- ---------- ---------- ---------- ---------- ---------- 2887 C*08:01:01 TTTCGAATGT GTGGTGAGCT GTTCAGAGTG TCATCACTTA CCATGACTGA CCTGAATTTG TTCATGACTA TTGTGTTCTG C*08:01:01NEW ---------- ---------- ---------- ---------- ---------- ---------- ---------- ---------- Exon8 2948 C*08:01:01 TAGCCTGAGA CAGCTGCCTG TGTGGGACTG AGATGCAGGA TTTCTTCACA CCTTTCCTTT G C*08:01:01NEW ---------- ---------- ---------- ---------- ---------- ---------- -

图1 标本1低分辨试剂盒扩增第2，3，4外显子后测序结果 Figure 1 Sample 1 sequencing results by low-resolution kit 图注：如图所示，标本1低分辨试剂盒扩增第2，3，4外显子得出的最接近结果为A*02∶03杂合A*11∶01，但有1个不匹配位点，疑似新基因。

图2 标本1低分辨试剂盒扩增第2，3，4外显子后测序结果 Figure 2 Sample 1 sequencing results by low-resolution kit 图注：如图所示，各孔均可见单一、清晰的约3 kb 电泳条带，其中M 为DL10 000 DNA marker ；1：MSA102(A*02∶03)；2：MSA111(A*11∶01)；3：MSC306(C*08∶01)；4：MSC308(C*07∶373)。

4 000 bp 3 000 bp

M 1 2 3 4

图3 新基因与A*11∶01∶01∶01 全长比对图

Figure 3 The new allele full-length comparison with HLA-A*11:01:01:01

图注：如图所示，该新基因与A*11∶01∶01∶01全长比较，在NT817由C 发生突变为T ，由于其发生在第三外显子上，故引起第125位密码子由GCC 变为GTC ，从而引起其编码的氨基酸由脯氨酸变成缬氨酸。

图4 标本2低分辨试剂盒扩增第2，3，4外显子后测序结果 Figure 4 Sample 2 sequencing results by low-resolution kit 图注：如图所示，标本2低分辨试剂盒扩增第2，3，4外显子得出的最接近结果为C*08∶01杂合C*07∶373，但C*07∶373在广东汉族人群中是一个相对罕见的等位基因；而图中右侧C*07∶02杂合C*08:01虽有一个错配碱基，但其为常见等位基因。

图5 新基因与C*08∶01∶01全长比对图

Figure 5 The new allele full-length comparison with HLA- C*08:01:01

图注：如图所示，该新基因与C*08∶01∶01比较，在NT879由A 发生突变为G ，由于其发生在第三外显子上，故引起第144位密码子由CAG 变为CGG ，从而引起其编码的氨基酸由谷氨酰胺变成精氨酸。