CAJ | 학술논문

目的筛查4例成骨不全(OI)家系先证者Ⅰ型前胶原α链基因(COL1A1/COL1A2)基因突变,分析基因型与表型的关系.方法收集先证者及家系成员临床资料,采集先证者、家系成员及50例正常对照的血液标本.采用聚合酶链反应(PCR)-高分辨率熔解曲线分析(HRMA)筛查先证者COL1A1/COL1 A2基因突变,基因测序确定突变位点.结果 HRMA显示先证者1分别在COL1A2基因12外显子、19外显子区域结果异常,标准熔解曲线与正常对照存在明显差异.测序结果,先证者1分别在COL1 A2基因12外显子、19外显子及18内含子发生突变:c.578G＞T、c.948 C＞T、c.937-3T＞C;其中c.948 C＞T、c.937-3T＞C为单核苷酸多态性(SNP)位点,c.578G＞T为新发现的突变位点.先证者2、3、4,分别在COL1 A2基因的38外显子、COL1A1基因的6外显子、23外显子区域的标准熔解曲线与正常对照存在明显差异.测序验证先证者2 COL1 A2基因突变:c.2305 G＞T.先证者3、4均在COL1A1基因发生突变:c.517 G＞T、c.1588 G＞A.其中先证者3 COL1A1基因c.517 G＞T为新发现的突变位点.先证者1、2、4基因突变,导致Ⅰ型胶原蛋白3股螺旋区域甘氨酸(Gly)被替换,可引起较为严重的表型,先证者3的无义突变,可造成Ⅰ型胶原合成量的改变,临床表型较轻.结论患者COL1A2基因c.578 G＞T、COL1A1基因c.517 G＞T均为新发现的突变位点,患者的基因型与临床表型存在一定的联系.
목적 사사4례성골불전(OI)가계선증자Ⅰ형전효원α련기인(COL1A1/COL1A2)기인돌변,분석기인형여표형적관계.방법 수집선증자급가계성원림상자료,채집선증자、가계성원급50례정상대조적혈액표본.채용취합매련반응(PCR)-고분변솔용해곡선분석(HRMA)사사선증자COL1A1/COL1 A2기인돌변,기인측서학정돌변위점.결과 HRMA현시선증자1분별재COL1A2기인12외현자、19외현자구역결과이상,표준용해곡선여정상대조존재명현차이.측서결과,선증자1분별재COL1 A2기인12외현자、19외현자급18내함자발생돌변:c.578G＞T、c.948 C＞T、c.937-3T＞C;기중c.948 C＞T、c.937-3T＞C위단핵감산다태성(SNP)위점,c.578G＞T위신발현적돌변위점.선증자2、3、4,분별재COL1 A2기인적38외현자、COL1A1기인적6외현자、23외현자구역적표준용해곡선여정상대조존재명현차이.측서험증선증자2 COL1 A2기인돌변:c.2305 G＞T.선증자3、4균재COL1A1기인발생돌변:c.517 G＞T、c.1588 G＞A.기중선증자3 COL1A1기인c.517 G＞T위신발현적돌변위점.선증자1、2、4기인돌변,도치Ⅰ형효원단백3고라선구역감안산(Gly)피체환,가인기교위엄중적표형,선증자3적무의돌변,가조성Ⅰ형효원합성량적개변,림상표형교경.결론 환자COL1A2기인c.578 G＞T、COL1A1기인c.517 G＞T균위신발현적돌변위점,환자적기인형여림상표형존재일정적련계.
Objective To investigate type Ⅰ collagen alpha chain gene (COL1A1 and COL1A2) gene mutation and analyze the correlation between genotype and clinical phenotype in four families with osteogenesis imperfecta (OI).Methods A survey was taken out of families history of OI probands along with the clinical data.We collected blood samples from the proband and their families,as well as 50 normal controls peripheral blood samples.Polymerase chain reaction analysis and high resolution melting (PCR-HRMA) method was used to screen blood samples COL1A1 and COL1A2 genes and validated by gene sequencing.Results PCR-HRMA method showed abnormal results from proband 1 in the COL1 A2 gene exon 12 and exon 19.Standard melting curves of proband 1 were differences from normal controls.Sequencing analysis showed that proband 1 with COL1A2 gene mutation,c.578G ＞ T,c.948 C ＞ T and c.937-3T ＞ C.That COL1A2 gene mutation c.948 C ＞ T and c.937-3T ＞ C were the SNP.The study found a new mutation of COL1A2 gene,c.578 G ＞T.PCR-HRM method showed abnormal results from proband 2,3 and 4 in the COL1 A2 gene exon 38,COL1 A1 gene exon 6 and exon 23,respectively.Standard melting curves of probands were differences from normal controls.Sequencing analysis showed that COL1A2 gene mutation c.2305G ＞ T,COL1A1 gene mutation c.517 G ＞ T,c.1588 G ＞ A,respectively.The study found a new mutation of COL1A1 gene,c.517 G ＞T.Genetic mutations from Proband 1,2 and 4 with OI in collagen type Ⅰ helix structure domain glycine (Gly) alternative,which cause a more severe clinical phenotype.Missense mutations in Proband 3 may result in type Ⅰ OI,which clinical phenotype is lighter.Conclusion COL1A2 gene mutation c.578G ＞ T,and COL1A1 gene mutation c.517 G ＞ T in patients with OI,are newly discovered mutation.Genotypes and clinical phenotypes in patients with OI may have some connection.