摘要目的：使用 Ion AmliSeq 多重 PCR 定制序列捕获系统及 Ion Torrent PGM 高通量测序平台对散发肌张力障碍患者进行肌张力障碍致病基因变异检测，以优化检测流程，探讨应用靶序列捕获结合高通量测序进行遗传病遗传诊断辅助临床干预的可能性。方法对95例确诊的肌张力障碍患者的外周血白细胞 DNA 使用 Ion AmliSeq 多重 PCR 进行靶序列捕获， Ion Torrent PGM 平台进行高通量测序，并对所得结果进行 Sanger 测序验证。结果本研究使用芯片12张，测序深度为134±44，覆盖度96.12％±0.71％。单张芯片测序周期为6小时（上机测序），单张芯片最高通量可达到900 Mb 以上。在95例肌张力患者中检出罕见变异71个，经 Sanger 验证准确率为41.04％，假阳性率58.96％。影响测序筛查质量主要原因在于 DNA 特殊序列如序列中存在同聚物。结论 Ion Torrent PGM 高通量测序平台的使用，具有测序通量高，操作简便，速度快，成本低的优点，可以提高基因突变筛查水平，在合理范围内扩增基因筛选数目，可以协助临床对肌张力障碍的鉴别诊断提供指导作用。

abstractsObjective To identity the pathogenic gene mutation in sporadic dystonia patients through Ion AmliSeq multiplex PCR custom sequence acquisition system and the Ion Torrent PGM high throughput sequencing platform , optimize the examination process , and explore the possibility of genetic di-agnosis and clinical intervention of genetic diseases through target sequence capture with high -throughput sequencing .Methods Totally 9 5 patients who were clinically diagnosed as dystonia in PUMCH were en-rolled .After extracting whole genome DNA from their peripheral blood leukocytes , we performed sequence by the Ion Torrent Personal Genome Machine ( PGM ) .Then we used Sanger sequence to validate all the variants .Results Totally 1 2 chips were used in the study , the sequencing depth was 1 3 4 ±4 4 and the coverage was 9 6.1 2 ±0.7 1%.The sequencing cycle of Single chip was 6 hours ( sequencing ) and the maximum data of single chip could reach 9 0 0 Mb .Among 9 5 patients with dystonia , 7 1 rare variants were detected .The accuracy rate was 4 1 .0 4% and the false positive rate was 5 8.9 6%.The main factor that influenced the quality of sequencing screening was the special sequence structure of DNA , such as the presence of the same polymer .Conclusion Ion Torrent PGM sequencing platform has the advantages of high sequencing throughput , easy operation , fast speed and low cost , which can improve the efficiency of genetic screening for genetic diseases .In a reasonable range of gene number , Ion Torrent PGM will help to provide guidance for the differential diagnosis of dystonia .