[应用高通量测序技术检测TSC1/TSC2基因突变用于结节性硬化症的快速诊断]
[Detection of TSC1/TSC2 gene mutation for rapid diagnosis of tuberous sclerosis complex by high-throughput sequencing technology].
作者信息
Huang Changyan
机构信息
Clinical laboratory, Ganzhou Municipal Hospital, Ganzhou 341000, China. *Corresponding author, E-mail:
出版信息
Xi Bao Yu Fen Zi Mian Yi Xue Za Zhi. 2016 Jan;32(1):92-5.
OBJECTIVE
To establish a high-throughput sequencing method for rapid detection of gene mutations of tuberous sclerosis complex (TSC1/TSC2) disease.
METHODS
A total of 10 patients with tuberous sclerosis disease and 10 healthy people were enrolled in this study. Long-chain polymerase chain reaction (PCR) was used to amplify all exon regions of TSC1 and TSC2 genes. The products were sequenced by Ion PGM(TM) platform and validated by Sanger sequencing.
RESULTS
All exons of TSC1 and TSC2 genes were specifically amplified and seven long segments were produced with the length between 13 000 bp to 15 000 bp. By Ion PGM machine, 10 pathogenic mutations were quickly identified. Among them, 7 were located in TSC1 and 3 were observed in TSC2. All variations were verified by Sanger sequencing.
CONCLUSION
Next generation sequencing (NGS) is an efficient method for rapid diagnosis of gene mutations of tuberous sclerosis complex.
目的
建立一种用于快速检测结节性硬化症复合物（TSC1/TSC2）疾病基因突变的高通量测序方法。
方法
本研究共纳入10例结节性硬化症患者和10名健康人。采用长链聚合酶链反应（PCR）扩增TSC1和TSC2基因的所有外显子区域。产物通过Ion PGM(TM)平台进行测序，并通过桑格测序进行验证。
结果
TSC1和TSC2基因的所有外显子均被特异性扩增，产生了7个长度在13 000 bp至15 000 bp之间的长片段。通过Ion PGM机器，快速鉴定出10个致病突变。其中，7个位于TSC1，3个在TSC2中被观察到。所有变异均通过桑格测序得到验证。
结论
下一代测序（NGS）是快速诊断结节性硬化症复合物基因突变的有效方法。
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