[Detection of TSC1/TSC2 gene mutation for rapid diagnosis of tuberous sclerosis complex by high-throughput sequencing technology].

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.