Application Of High-through Put Sequencing Platform In Genetic Analysis Of Dystonia

Background Dystonia is a movement disorder characterized by sustained or intermittent muscle contractions causing abnormal, often repetitive, movements, postures, or both. It may results in significant disability. Due to the clinical and genetic heterogeneity of dystonia, a rapid, accurate method with parallel screening capacity of multiple genes would profoundly improve its diagnostic status.Object To apply Ion AmliSeq multiple PCR custom sequence capture system combined with Ion Torrent PGM high-through put sequencing platform in the mutation analysis of patients with dystonia. And to evaluate the effectiveness of high throughput sequencing technology in clinical diagnosis of dystonia.Methods The custom panel includes all the exons and their flanking sequences of 25 dystonia genes and other dystonia-associated genes. A panel with ion AmliSeq multiplex PCR custom sequence capture system was apply to capture the coding sequence of the targeted genes. Then the ion torrent PGM high-throughput sequencing platform was used to generate the sequencing data. Finally the resultant variants were analyzed through the pipeline.Result In this study, we used 6 Ion Chip Ion 318 chips and 12 Ion Chip 318V2 chips, the average depth of sequencing was 134+23,96.12%+0.71% of target zone was covered with the maximum generating data of single chip reaching 900Mb (Ion Chip 318V2 chip). After the data analysis and verification by Sanger sequencing,15(60%, 15/25) mutations were detected in the causing genes of dystonia. Among them, there are 4 known mutations, and the other 11 variants have not been reported before.Conclusion Application of Ion AmliSeq multiple PCR custom sequence capture system combined with Ion Torrent PGM high-throughput sequencing platform can find pathogenic mutations in patients with dystonia rapidly and help to diagnose the disease precisely.