These tools will no longer be maintained as of December 31, 2024. Archived website can be found here. PubMed4Hh GitHub repository can be found here. Contact NLM Customer Service if you have questions.


Pubmed for Handhelds

PUBMED FOR HANDHELDS

Search MEDLINE/PubMed

  • Title: [Sanger sequencing for the diagnosis of spinal muscular atrophy patients with survival motor neuron gene 1 compound heterozygous mutation].
    Author: Yang L, Cao YY, Qu YJ, Bai JL, Wang H, Jin YW, Han YL, Song F.
    Journal: Zhonghua Yi Xue Za Zhi; 2017 Feb 14; 97(6):418-423. PubMed ID: 28219127.
    Abstract:
    Objective: To detect the subtle variant of survival motor neuron gene 1(SMN1) by Sanger sequencing, and to assess the value of Sanger sequencing for the diagnosis of spinal muscular atrophy(SMA) with compound heterozygous mutation of SMN1. Methods: Fifty-two patients suspected SMA were recruited by the Capital Institute of Pediatrics from Jan.2014 to June.2016. PCR was used for amplifying exon7 of SMN1 and SMN2 in 52 patients. Natural different base peaks on the sequencing chromatogram in the SMN1 and SMN2 within the amplified segments were identified with Sanger DNA sequencing to detect the homozygous deletion or heterozygous deletion of SMN1. Then we screened the SMN1 subtle variants in heterozygous deletion patients by genomic Sanger sequencing for the other SMN exons. At last, multiplex ligation-dependent probe amplification(MLPA) was carried out to confirm the results of SMN1 heterozygous deletion, and T-A cloning confirmed the subtle variants were located in SMN1. Results: Forty-seven of 52 cases were homozygous deletion of SMN1, while 5 cases were heterozygous deletion which were confirmed by MLPA.Then, by genomic and T-A cloning sequencing, five SMN1 subtle mutations were separately identified in 5 cases of heterozygous deletion. Conclusion: Sanger sequencing is an effective method for the clinical diagnosis of compound heterozygous mutation of SMN1, and is meaningful for improving genetic diagnosis rate of SMA. 目的：建立并评价Sanger测序技术在诊断运动神经元存活基因1(SMN1)复合杂合突变型脊髓性肌萎缩症(SMA)病例中的意义。 方法：选取首都儿科研究所2014年1月至2016年6月就诊的52例SMA患儿，应用聚合酶链反应(PCR)技术扩增52例患儿的SMN1和SMN2基因第7外显子，Sanger测序通过识别扩增片段中SMN1和SMN2基因固有差异碱基的峰高，判断SMN1基因纯合或杂合缺失。当SMN1基因杂合缺失时，经PCR-Sanger测序筛查SMN基因所有外显子、外显子与内含子交界区域的变异位点。最后通过多重连接探针扩增(MLPA)方法验证SMN1基因杂合缺失，并利用克隆测序确定突变点来自SMN1基因，完成SMN1基因复合杂合突变的确诊。 结果：经Sanger测序检测的52例SMA患儿中47例为SMN1基因纯合缺失，5例为SMN1基因杂合缺失。MLPA方法验证了后5例患儿SMN1基因拷贝数均为1，Sanger测序检测出5例患儿均存在SMN基因点突变，并通过克隆Sanger测序验证了5例患儿突变均位于SMN1基因。 结论： Sanger测序技术适用于SMN1基因复合杂合突变病例的诊断，对提高SMA基因诊断率具有临床意义。.
    [Abstract] [Full Text] [Related] [New Search]