上海交通大学学报（医学版）

上海交通大学学报（医学版）

• 论著（基础研究） • 上一篇    下一篇

嗜铬细胞瘤7个致病基因的高通量基因检测方法的建立

谢晓雁1，宋倩倩1，崔 斌1,2   

  1. 1. 上海交通大学 医学院附属瑞金医院，上海市内分泌代谢病研究所，上海 200025；2. 中国科学院上海生命科学研究院/上海交通大学医学院 健康科学研究所，上海 200025
  • 出版日期:2016-08-29 发布日期:2016-08-31
  • 通讯作者: 崔 斌，电子信箱：bcui@sibs.ac.cn。
  • 作者简介:谢晓雁（1972—），女，主管技师，硕士；电子信箱：xiaoyan115@hotmail.com。
  • 基金资助:

    上海市浦江人才计划（15PJD029）；上海市教育委员会高峰高原学科建设计划（20152502）

Establishment of a high throughput gene detection method for seven pathogenic genes of pheochromocytoma

XIE Xiao-yan1, SONG Qian-qian1, CUI Bin1,2   

  1. 1. Shanghai Institute of Endocrinology and Metabolism, Ruijin Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai 200025, China; 2. Institute of Health Sciences, Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences / Shanghai Jiao Tong University School of Medicine, Shanghai 200025, China
  • Online:2016-08-29 Published:2016-08-31
  • Supported by:

    Shanghai Pujiang Program, 15PJD027; Shanghai Municipal Education Commission—Gaofeng Clinical Medicine Grant Support, 20152502

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摘要：

目的·建立嗜铬细胞瘤致病基因的高通量测序检测方法平台体系，为基因诊断进入常规临床检测作准备。方法·采用高通量测序建库两步PCR方法，针对嗜铬细胞瘤的7个致病基因（SDHB, SDHC, SDHD, VHL, MAX, TMEM127和RET）的35个基因片段进行建库，然后使用Miseq机器进行后续的高通量测序。结果·建库的35个基因片段在Miseq机器都得到了检测，并且均一性达到了实验最初的设想。结论·该实验方法完成了嗜铬细胞瘤7个致病基因的基因检测工作，实验的质量和成本都达到了预期效果。

关键词: 嗜铬细胞, 高通量测序, 致病基因

Abstract:

Objective · To establish a test method platform system for high throughput sequencing of pathogenic genes of pheochromocytoma and make preparation for including the genetic diagnosis into routine clinical testing. Methods · The bank was constructed for 35 DNA fragments in 7 pathogenic genes (SDHB, SDHC, SDHD, VHL, MAX, TMEM127, and RET) of pheochromocytoma with two-step PCR method of high throughput sequencing. The high throughput sequencing was performed on illumine Miseq machine. Results · Thirty-five DNA fragments were detected on illumine Miseq machine and the uniformity met the design of experiment. Conclusion · This method completed the gene detection for 7 pathogenic genes of pheochromocytoma. The quality and cost of the experiment achieved the expectation.