中华医学遗传学杂志
中華醫學遺傳學雜誌
중화의학유전학잡지
CHINESE JOURNAL OF MEDICAL GENETICS
2015年
1期
40-43
,共4页
和艳敏%陶苏丹%章伟%王炜%何吉%朱发明%吕杭军
和豔敏%陶囌丹%章偉%王煒%何吉%硃髮明%呂杭軍
화염민%도소단%장위%왕위%하길%주발명%려항군
HLA-DPB1基因%第3外显子%多态性%造血干细胞移植
HLA-DPB1基因%第3外顯子%多態性%造血榦細胞移植
HLA-DPB1기인%제3외현자%다태성%조혈간세포이식
HLA-DPB1 gene%Exon 3%Polymorphism%Hematopoietic stem cell transplantation
目的 建立HLA-DPB1基因第2～3外显子测序方法,并分析其多态性.方法 根据HLA-DPB1基因序列,设计位点特异性引物扩增242名浙江汉族人群HLA-DPB1第2～3外显子序列,PCR扩增产物经双酶切后对第2和3外显子进行双向测序分析,采用Assign 3.5+SBT分析软件判读HLA-DPB1等位基因型.结果 PCR扩增获得特异性的单一目的片段,其产物经直接测序后得到清晰的序列图.242名浙江汉族人群中共检测到18个HLA-DPB1等位基因,频率超过0.05的等位基因为DPB1*05:01:01/135:01(0.4112)、DPB1*02:01:02(0.1901)、DPB1*04:01:01(0.1136)以及DPB1 *02:02(0.0620),并确认1个新等位基因DPB1*168:01.在第3外显子中发现9个多态性位点,其中517 A＞T为本实验新检出的1个多态性位点.结论 本实验建立的HLA-DPB1第2～3外显子测序方法是可行的,HLA-DPB1第3外显子存在一定多态性.
目的 建立HLA-DPB1基因第2～3外顯子測序方法,併分析其多態性.方法 根據HLA-DPB1基因序列,設計位點特異性引物擴增242名浙江漢族人群HLA-DPB1第2～3外顯子序列,PCR擴增產物經雙酶切後對第2和3外顯子進行雙嚮測序分析,採用Assign 3.5+SBT分析軟件判讀HLA-DPB1等位基因型.結果 PCR擴增穫得特異性的單一目的片段,其產物經直接測序後得到清晰的序列圖.242名浙江漢族人群中共檢測到18箇HLA-DPB1等位基因,頻率超過0.05的等位基因為DPB1*05:01:01/135:01(0.4112)、DPB1*02:01:02(0.1901)、DPB1*04:01:01(0.1136)以及DPB1 *02:02(0.0620),併確認1箇新等位基因DPB1*168:01.在第3外顯子中髮現9箇多態性位點,其中517 A＞T為本實驗新檢齣的1箇多態性位點.結論 本實驗建立的HLA-DPB1第2～3外顯子測序方法是可行的,HLA-DPB1第3外顯子存在一定多態性.
목적 건립HLA-DPB1기인제2～3외현자측서방법,병분석기다태성.방법 근거HLA-DPB1기인서렬,설계위점특이성인물확증242명절강한족인군HLA-DPB1제2～3외현자서렬,PCR확증산물경쌍매절후대제2화3외현자진행쌍향측서분석,채용Assign 3.5+SBT분석연건판독HLA-DPB1등위기인형.결과 PCR확증획득특이성적단일목적편단,기산물경직접측서후득도청석적서렬도.242명절강한족인군중공검측도18개HLA-DPB1등위기인,빈솔초과0.05적등위기인위DPB1*05:01:01/135:01(0.4112)、DPB1*02:01:02(0.1901)、DPB1*04:01:01(0.1136)이급DPB1 *02:02(0.0620),병학인1개신등위기인DPB1*168:01.재제3외현자중발현9개다태성위점,기중517 A＞T위본실험신검출적1개다태성위점.결론 본실험건립적HLA-DPB1제2～3외현자측서방법시가행적,HLA-DPB1제3외현자존재일정다태성.
Objective To establish a polymerase chain reaction sequencing-based typing (PCR-SBT) method for HLA-DPB1 exons 2 and 3,and to analyze their polymorphisms.Methods Based on the sequences of HLA-DPB1 loci,locus-specific primers were designed and applied to amplify the target sequences encompassing the entire exons 2 and 3 of HLA-DPB1.The amplification products were digested by enzymes and directly sequenced in both directions.The genotype was assigned by Assign 3.5 + SBT software.Results Specific target fragment was obtained with the PCR amplification,and good quality electropherogram was derived by direct sequencing.Among 242 individuals from Zhejiang Han population,18 HLA-DPB1 alleles were detected.Alleles with a frequency of＞0.05 have included DPB1 * 05:01:01/135:01 (0.4112),DPB1 *02:01:02 (0.1901),DPB1 *04:01:01 (0.1136) andDPB1 *02:02 (0.0620).A novel HLA-DPB1 * 168:01 allele has also been identified.Nine polymorphism sites were founded in the exon 3 region,which included a new SNP site 517 A＞T.Conclusion The PCR-SBT method for exons 2 and 3 of HLA-DPB1 is reliable,which allowed detection of polymorphisms in exon 3 of the HLA-DPB1 gene.
