CAJ | 학술논문

目的建立一套运动神经元生存(SMN)基因微小突变检测体系,以评价其用于脊肌萎缩症(SMA)家系的价值.方法采用RNA水平和基因组DNA水平双途径检测策略,用PCR-限制性片段长度多态性(RFLP)、位点特异性PCR、多重连接依赖性探针扩增(MLPA)和T克隆测序技术对2个SMA家系共7个成员行SMN基因微小突变分析.结果用MLPA和T克隆测序技术检测家系A的SMA患者有1个拷贝的SMN1基因,其第228位密码子上存在1个无义突变L228X,该突变来源于患者父亲；家系B的SMA患者父亲有2个SMN1基因,其中1个SMN1基因存在移码突变22_23 insA.其余家系成员均为有1个SMN1基因拷贝的SMA携带者.结论建立的SMN微小突变检测体系成功鉴定了2个SMN微小突变,为SMA家系的遗传咨询提供了可靠依据.
목적 건립일투운동신경원생존(SMN)기인미소돌변검측체계,이평개기용우척기위축증(SMA)가계적개치.방법 채용RNA수평화기인조DNA수평쌍도경검측책략,용PCR-한제성편단장도다태성(RFLP)、위점특이성PCR、다중련접의뢰성탐침확증(MLPA)화T극륭측서기술대2개SMA가계공7개성원행SMN기인미소돌변분석.결과 용MLPA화T극륭측서기술검측가계A적SMA환자유1개고패적SMN1기인,기제228위밀마자상존재1개무의돌변L228X,해돌변래원우환자부친；가계B적SMA환자부친유2개SMN1기인,기중1개SMN1기인존재이마돌변22_23 insA.기여가계성원균위유1개SMN1기인고패적SMA휴대자.결론 건립적SMN미소돌변검측체계성공감정료2개SMN미소돌변,위SMA가계적유전자순제공료가고의거.
Objective To establish a analytical system for the survival motor neuron (SMN) subtle mutation,and evaluate its application in two families with spinal muscular atrophy (SMA).Methods SMN genes in seven family members from two SMA families were analyzed at both transcript level and genomic level,by the use of the conventional PCR-RFLP,allele-specific PCR,multiplex ligation-dependent probe amplification (MLPA) and T subcloning and sequencing of SMNI gene.Results In family A,the patient had a single SMN1 copy who was carrying nonsense mutation L228X,which was also found in his father.In family B,as the patient's sample was unavailable,the father was indeed a carrier with one normal SMN1 allele and the other SMN1 allele carrying a frameshift mutation 22_23insA.The remaining family members were SMA carriers with one SMN1 copy.Conclusion This analytical system for SMN subtle mutation offers viable molecular basis for genetic counseling and prenatal diagnosis in SMA families.