摘要
目的应用PCR-RFLP和多重连接依赖式探针扩增法(MLPA)技术对临床诊断为脊髓性肌萎缩症(SMA)患儿进行基因诊断,并比较两种方法检测SMN基因缺失改变的效果。方法应用DNA抽提试剂盒抽提3个SMA家系成员的外周血样品DNA,电泳检测,定量,常规PCR法扩增SMN基因的7、8号外显子,Dra I、Dde I酶切PCR产物,常规聚丙烯胺凝胶电泳法分离酶切后PCR产物,Genefinder染色检测。同时应用M LPA检测试剂盒P021进行检测。结果 PCR-RFLP显示3个家系中的患儿均纯合缺失SMN1的第7、8号外显子,MLPA检测结果与之完全相符。结论与PCR-RFLP相比,MLPA更加简便、快捷、可靠,是一种高效的遗传病基因诊断手段。
Objective：To perform gene diagnosis for spinal muscular atrophy（SMA） using PCR-restriction fragment length polymorphism（PCR-RFLP） and multiplex ligation dependent probe amplification（MLPA）,and compare the effects of the two methods on detecting SMN gene deletion. Methods ：Genomic DNA was isolated from peripheral blood of each subject using DNA Purification kit method. DNA concentration was determined and quantified by electrophores.Exon7and8 of the SMN gene was amplified by allele specific PCR. The PCR products were digested with Dra I,Dde I separated by PAGE（polyacrylamide gel electrophoresis）,and visualized by Genefinder staining. Simultaneously,the DNA samples were analyzed applying SALSA MLPA K IT P〈021. Results ：Both PCR-RFLP and M LPA analysis showed that all the 3 patients with SMApresentedexon7and8 homozygous deletion of SMN1.Conclusion：Compared with PCR-RFLP,MLPA is more convenient,high-effective and reliable. It is a conventional technique of gene diagnosis for common congenital genetic disease with high incidence.
作者
闫杨
王红梅
宋楠
黄胜男
孙斌
YAN Yang WANG Hong-mei SONG Nan HUANG Sheng-nan SUN Bing.(The 463 Hospital of PLA, Cyto-therapy Center, Liaoning Shenyang, 110042)
出处
《中国优生与遗传杂志》
2017年第4期49-51,共3页
Chinese Journal of Birth Health & Heredity
