[Sanger sequencing for the diagnosis of spinal muscular atrophy patients with survival motor neuron gene 1 compound heterozygous mutation].
Zhonghua Yi Xue Za Zhi
; 97(6): 418-423, 2017 Feb 14.
Article
em Zh
| MEDLINE
| ID: mdl-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.Palavras-chave
Texto completo:
1
Coleções:
01-internacional
Base de dados:
MEDLINE
Assunto principal:
Atrofia Muscular Espinal
Tipo de estudo:
Diagnostic_studies
Limite:
Humans
Idioma:
Zh
Revista:
Zhonghua Yi Xue Za Zhi
Ano de publicação:
2017
Tipo de documento:
Article
País de afiliação:
China