一个成骨不全家系COL1A1基因的突变筛查.pdf

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1、中华医学遗传学杂志2014年6月第31鲞第3盟hi!：!!：!!：临床遗传学论著一个成骨不全家系COLIA1基因的突变筛查白雪李克秋任秀智何晓波王毅官士珍景亚青李光【摘要】目的筛查1个成骨不全(osteogenesisimperfecta，OI)家系中COL1A1基因的突变，并分析基因型与临床表型的关系。方法收集先证者及家系成员临床资料，采集先证者、随诊家属及5O名正常对照的外周血标本，应用PCR一高分辨率熔解曲线(highresolutionmelting，HRM)分析筛查COL1A1基因突变，基因测

2、序确定突变位点。结果PCR-HRM分析显示，先证者COLIA1基因第33～34外显子结果异常，Tm值为87．7℃，比正常对照的Tm值(87．9℃±0．06℃)低0．2℃，先证者与正常对照标准熔解曲线及差异熔解曲线均有明显差异。测序结果提示，先证者COL1A1基因第33～34外显子c．232ldelC，移码突变，导致此后第334位氨基酸提前出现终止密码子(UAA)，即P．Pro774IeufsX334杂合突变。先证者之父、祖父基因测序均具有相同的突变位点。5o名正常对照无此突变。该突变位点在人类胶原突变数

3、据库中未见报道。家系图谱显示为常染色体显性遗传，先证者及家系患者临床诊断均为I型0I。结论成骨不全C0L】A1基因c．2321delC为新的突变位点。移码突变导致提前形成终止密码子，使I型胶原合成量减少，临床表型较轻。【关键词】成骨不全；C0LjA】基因；基因突变；基因诊断；高分辨率熔解曲线ScreeningandanalysisofanewmutationofCOL1A1geneinafamilywithosteogenesisimperfectaBaiXue，LiKeqiu，RenXiuzhi。，H

4、eXiaobo，WangYi，GuanShizhen，JingYaqing，LiGuang。．(DepartmentofMedicalLaboratory，TianjinHospital，Tianjin300211，P．R．China)；(DepartmentofBiology，TianjinMedicalUniversity，Tianjin300070，P．R．China)；。(DepartmentofPediatricOrthopedics，TianjinHospital，Tianjin300211

5、，P．R．China)C0rrs户ongauthor：LiGuang，Email：lig@tijmu．edu．cn[Abstract]ObjectiveToinvestigatemutationofCOL1A1geneandanalyzetherelationshipbetweengenotypeandclinicalphenotypeinafamilywithosteogenesisirnperfecta(0I)．MethodsThefamilyhistoryofanOIpedigree，alongw

6、ithclinicaldata，wascollected．Bloodsamplesfromtheprobandandhisfamilies，aswellas50normalcontrols，werecollected．MutationofCOLIA1genewasscreenedusingPCR—highresolutionmelting(PCR—HRM)andvalidatedbysequencing．ResultsPCR—HRMmethodshowedanabnorma1resultinproban

7、dC0L1A13334exons，whichTmwas87．7℃，incontrasttothenormalcontro1—(wt)Tmof87．9C土0．06℃．Therewasasignificantdifferencebetweentheprobandandthenorma1controlwiththestandardizationcurveandthedifferencecurves．DNAsequencingshowedthatCOL1A1geneexons33—34has1ostaCbase

8、(c．2321delC)，whichresultedinaframeshiftmutationandcausedanprematureterminationcodon(UAA)ataminoacid334，i．e．，P．Pro774LeufsX334．Thefatherandgrandfatheroftheproband，bothsufferedfromOI，wereverifiedtobeheterozygousforthesamemut