摘要
目的进行脊髓性肌萎缩症(spinal muscular atrophy,SMA)基因携带者的筛查,为遗传咨询提供理论依据。方法应用实时荧光定量PCR特异性扩增264名健康人、88例经基因诊断确诊SMA患者的双亲、32名SMA家系其它成员的SMN1基因第7外显子及其邻近区域,以已确定只有2拷贝SMN1的样品作为标准对照。结果88例确诊SMA患者双亲除4名SMN1拷贝数为2外,其余均只有1拷贝SMN1。264名正常人中5人仅有1拷贝SMN1,为基因携带者,该组中含2、3、4拷贝SMN1的人数分别为232、25、2。32名SMA家系成员中有2名SMN1拷贝数为1,为基因携带者,25名SMN1拷贝数为2,另5名拷贝数为3。结论实时荧光定量PCR技术可进行单拷贝差异SMN1基因的定量检测,结果准确、重复性好,基因携带者的筛查为本病遗传咨询提供了重要依据。
Objective To construcl a method for detecting the copy number of survival of motor meuron I gene （SMNI）with single copy difference based on real-time fluorescence quantitative PCR.and to make practical use of the method for acquiring the data on SMN1 copy number in Chinese as well as for screening the carriers of spinal muscular atrophy（SMA） from bealthy individuals and SMA families.Methods：Exon 7 and flanking area of SMN1 gene were amplified by real-time fluoreseence quantitative PCR in 264 healthy individuals.in 1 standand sample having 2 SMN1 but having no SMN2,and in 88 parents of SMA patients.The samples for detecting were diluted to 30ng/μL and the standard sample was diluted to 15ng/μL.30ng/μL,45ng/μL,60ng/μL;the nuknown samples and 4 standard samples with different concentrations were amplified at the same time,a standand curve coule be drawn out according to the results of the 4 standard samples,then the copy number of samples coule be calculated.Results Of 88 parents＇ samples.84 samples each had I copy of SMN1,and the rest 4 each had 2 copies of SMN1,Of 264 healthy individuals＇ samples,5 samples each had only I copy of SMN1（an indicator of definite gene carriers）.232 samples each had 2 copies of SMN1,25 samples each had 3 copies of SMN1,and 2 samples each had 4 copies of SMN1.Of the samples of 32 members of SMA families,2 samples each had only 1 copy of SMN1 indicating definite gene carriers,25 samples each had 2 copies of SMN1,and 5 samples each had 3 copies of SMN1.Conclusion：SMN1 copy number could be de tected precisely by real-time fluoreseence quantitative PCR;the sereening of gene carriers could provide essential data for genetie counseling.
出处
《中华医学遗传学杂志》
CAS
CSCD
北大核心
2005年第6期599-602,共4页
Chinese Journal of Medical Genetics
基金
福建省重大科技项目(2002Y001)~~
