CAJ | 학술논문

目的探讨2例类孟买血型个体的分子遗传机制.方法先证者为女性,在无偿献血时发现ABO血型正反定型不符,遂将其本人及家系成员(包括先证者爷爷、奶奶、父亲、母亲、弟弟、妹妹)血液标本(EDTA抗凝)和唾液标本送至广州血液中心进一步鉴定.采用常规血清学方法对先证者及其家系成员血液标本进行血型鉴定,对唾液标本进行ABH血型物质的检测；利用PCR扩增先证者及家系成员的FUT1和FUT2基因编码区和ABO血型基因第6、7外显子编码区,对PCR产物进行直接测序后分析结果,对FUTI基因的缺失突变进行克隆测序分析.结果先证者及其弟弟为类孟买血型,其他家系成员中未发现类孟买血型；直接测序结果显示先证者和弟弟的FUT1基因为第547 -552位碱基AG缺失、880-882位碱基TT缺失的杂合型；其爷爷和父亲为单链880-882位碱基TT缺失杂合型,母亲和妹妹为单链547-552位碱基AG缺失杂合型；克隆测序结果证实上述缺失分别发生在FUT1基因编码区第547-548位和第881-882位；先证者和弟弟的FUT2基因编码区均存在第390位C＞T、第749位G＞A突变,家系成员中还存在第418位A＞T突变.结论 FUT1基因不同位点双碱基缺失杂合可导致类孟买血型,单链缺失突变杂合型的ABO表型正常；发现了FUT2基因中3个新的突变位点.
목적 탐토2례류맹매혈형개체적분자유전궤제.방법 선증자위녀성,재무상헌혈시발현ABO혈형정반정형불부,수장기본인급가계성원(포괄선증자야야、내내、부친、모친、제제、매매)혈액표본(EDTA항응)화타액표본송지엄주혈액중심진일보감정.채용상규혈청학방법대선증자급기가계성원혈액표본진행혈형감정,대타액표본진행ABH혈형물질적검측；이용PCR확증선증자급가계성원적FUT1화FUT2기인편마구화ABO혈형기인제6、7외현자편마구,대PCR산물진행직접측서후분석결과,대FUTI기인적결실돌변진행극륭측서분석.결과 선증자급기제제위류맹매혈형,기타가계성원중미발현류맹매혈형；직접측서결과현시선증자화제제적FUT1기인위제547 -552위감기AG결실、880-882위감기TT결실적잡합형；기야야화부친위단련880-882위감기TT결실잡합형,모친화매매위단련547-552위감기AG결실잡합형；극륭측서결과증실상술결실분별발생재FUT1기인편마구제547-548위화제881-882위；선증자화제제적FUT2기인편마구균존재제390위C＞T、제749위G＞A돌변,가계성원중환존재제418위A＞T돌변.결론 FUT1기인불동위점쌍감기결실잡합가도치류맹매혈형,단련결실돌변잡합형적ABO표형정상；발현료FUT2기인중3개신적돌변위점.
Objective To study the molecular genetic mechanism of para-bombay phenotype in two individuals.Methods The proband was a female.When the proband donated blood,because the forward blood group wasn't coincident with her reverse blood group,the blood and saliva specimen from proband and her family members were sent to Guangzhou Blood Center for further identification.Routine serological techniques were used to determine proband's and her family members' blood group and ABH antigen in saliva.The coding regions of FUT1 and FUT2 gene,exon 6 and exon 7 of ABO gene were amplified by polymerase chain reaction using proband's and her family members' genomic DNA.All amplified products were analyzed after being directly sequenced.The two-base deletion regions of FUT1 gene were certified by cloning and haplotype sequencing.Results Proband's and her little brother's blood group were identified as para-bombay while other family members' blood group were normal.Two-base deletion heterozygous mutations of FUT1 gene were found in proband and her brother,AG deletion at position 547-552 and TT deletion at position 880-882,which caused a reading frame shift and a premature stop eodon.Meanwhile,880-882del TT heterozygous mutation was found in proband's grandfather and her father and 547-552del AG heterozygous mutation was found in proband's mother and her little sister.Results Of cloning and haplotype sequencing certified that these two-base deletion mutations occurred at 547-548 and 881-882 position respectively.Three new mutations were found in FUT2 gene,390C ＞ T,418A ＞ T and 749G ＞ A,which could cause the change of amino acid at position 140Ile ＞ Phe and 250Arg ＞ Gln.Conclusions Two-base deletion heterozygous mutations in different positions in FUT1 gene were found in 2 individuals,which maybe the molecular genetic mechanism of para-bombay phenotype.Heterozygous deletion mutation in one-strand DNA wouldn't change the ABO blood group.Three new mutations were also found in FUT2 gene.( Chin J Lab Med,2012,35:815-819)