CAJ | 학술논문

背景:目前所发现的HLA等位基因是在人类进化过程中,为了适应某种外界环境过程中逐渐产生的,新近产生而不曾被人们发现的为新等位基因,这些新的等位基因产生诱因千变万化,在诱发因素消除之后这些突变是否持续存在,尚需进一步观察.目的:采用DNA测序技术确认聚合酶链反应-序列特异性寡核苷酸分型检测中的可疑结果,确认新的HLA等位基因,分析新等位基因HLA-A~*9217(WHO注册号:WHS10004629)的遗传方式.设计、时间及地点:以DNA为观察对象的开放性试验.其中常规初检聚合酶链反应-寡核苷酸探针分型反向杂交荧光微珠法于2007-11在河南省红十字血液中心中华骨髓库河南分库人类白细胞抗原组织配型实验室完成,测序实验于2008-02在戴诺生物技术(北京)有限公司人类白细胞抗原实验室完成.材料:中华骨髓库志愿捐献者(编号:371xxxxx)及其直系3代亲属血样,受试者共9人在实验室逐个登记,采集静脉血5 mL,EDTA抗凝.方法:用聚合酶链反应-序列特异性寡核苷酸流式荧光微珠HLA分型方法、SBT测序分型方法,对A~*9217携带者家庭成员进行中低分辨,DNA测序高分辨检测分析,血清学方法检测ABO,MN,Rh血型系统,分析新基因的发现及遗传方式.主要观察指标:HLA新等位基因A~*9217携带者A位点外显子3序列对比.结果:通过受检者HLA分型结果和红细胞系统ABO,MN,Rh血型分析,先证者A~*9217应来自父方,但父方并没有检出A~*9217,而是检出了与之序列相近的A~*020301,与A~*9217在处显子3有3处碱基改变391 T>G,414 C>G,418 T >G,A~*9217携带者的2个孩子均检出A~*9217.结论:新基因A~*9217先证者父方产生突变所致,该突变能正常遗传给后代,该基因是否能稳定遗传下去有待进一步观察研究.
배경:목전소발현적HLA등위기인시재인류진화과정중,위료괄응모충외계배경과정중축점산생적,신근산생이불증피인문발현적위신등위기인,저사신적등위기인산생유인천변만화,재유발인소소제지후저사돌변시부지속존재,상수진일보관찰.목적:채용DNA측서기술학인취합매련반응-서렬특이성과핵감산분형검측중적가의결과,학인신적HLA등위기인,분석신등위기인HLA-A~*9217(WHO주책호:WHS10004629)적유전방식.설계、시간급지점:이DNA위관찰대상적개방성시험.기중상규초검취합매련반응-과핵감산탐침분형반향잡교형광미주법우2007-11재하남성홍십자혈액중심중화골수고하남분고인류백세포항원조직배형실험실완성,측서실험우2008-02재대낙생물기술(북경)유한공사인류백세포항원실험실완성.재료:중화골수고지원연헌자(편호:371xxxxx)급기직계3대친속혈양,수시자공9인재실험실축개등기,채집정맥혈5 mL,EDTA항응.방법:용취합매련반응-서렬특이성과핵감산류식형광미주HLA분형방법、SBT측서분형방법,대A~*9217휴대자가정성원진행중저분변,DNA측서고분변검측분석,혈청학방법검측ABO,MN,Rh혈형계통,분석신기인적발현급유전방식.주요관찰지표:HLA신등위기인A~*9217휴대자A위점외현자3서렬대비.결과:통과수검자HLA분형결과화홍세포계통ABO,MN,Rh혈형분석,선증자A~*9217응래자부방,단부방병몰유검출A~*9217,이시검출료여지서렬상근적A~*020301,여A~*9217재처현자3유3처감기개변391 T>G,414 C>G,418 T >G,A~*9217휴대자적2개해자균검출A~*9217.결론:신기인A~*9217선증자부방산생돌변소치,해돌변능정상유전급후대,해기인시부능은정유전하거유대진일보관찰연구.
BACKGROUND: The human leucocyte antigen (HLA) allele are now understood to be alternative DNA sequences at the same physical gene locus, which may or may not result in different phenotypic traits. The generation of a new allele is induced by various factors, but, whether the mutation would be exist after removing the causative factors need further investigation. OBJECTIVE: To confirm the new allele by DNA sequencing technique, in addition, to analyze the heritage of a HLA allele A~*9217 (No. of WHO registration: WHS10004629)DESIGN, TIME AND SETTING: The open experiment with DNA as observation object. The initial detection with polymerase chain reaction sequence-specific oligonucleotide (PCR-SSO) was performed at HLA Laboratory of Henan Provincial Red Cross Blood Center in November 2007, and the sequencing was performed at HLA Laboratory of DYNAL Biological Technology (Beijing) Co., Ltd. in February, 2008. MATERIALS: The proband (sample ID: 371xxxxx ) and other 8 family member was investigated, blood sample was collected at the HLA Laboratory.METHODS: Nine family members of A~*9217 carrier were typing for HLA- A, B, DRB1 using PCR-SSO and SBT methods for low-media and high resolution, and 3 red blood cell blood groups systems: ABO, MN and Rh was tested to assist analysis .MAIN OUTCOME MEASURES: Sequence alignment of HLA-A exon 3 in A~*9217 carriers.RESULTS: According to the results of the blood groups phonotype of ABO, MN, Rh systems and HLA, the new allele A~*9217 of the proband was paternal origin. However, the sequence of this allele is A*020301, which had 3 base difference in exon 3, the new allele appears 3 base change at 391 T>G, 414 C>G, 418 T>G, and this new sequence was found in proband's two children.CONCLUSION: The new allele A~*9217 is generated from proband's father mutation, which can pass it to his children. However, the further heritage of A~*9217 allele need exploring.