Ion Torrent PGM测序技术在MPN患者临床诊断中的方法学建立  

Establishing the Ion Torrent PGM Sequencing Methods for the Clinical Diagnosis of MPN Patients

作　　者：黄继贤[1,2] 李玉玲[1] 许娜[1] 阴常欣[1] 周璇[1] 潘成云[1] 何柏林[1] 陆紫媛 刘启发[1] 刘晓力[1] 

机构地区：[1]南方医科大学南方医院血液科,广东广州510515 [2]粤北人民医院血液内科,广东韶关512025

出　　处：《中国实验血液学杂志》2017年第6期1744-1750,共7页Journal of Experimental Hematology

基　　金：国家自然科学基金青年项目(81700104);广东省医学科研基金立项项目(A2017543);韶关市科技计划项目(2017cx/008);韶关市卫生计生科研项目(Y17019)

摘　　要：目的:探讨通过自主设计定制的MPN致病相关基因的多重PCR引物试剂盒,使用Ion Torrent PGM第二代测序平台快速和准确地获取MPN患者基因突变信息的可行性和可靠性。方法:收集2015年1月至2015年10月就诊于南方医院血液科,经Sanger测序法检测JAK2V617F+和(或)CALR+、Ph-的10例MPN患者骨髓标本,然后使用Ion Torrent PGM第二代测序复核检测骨髓并对比两种方法结果的一致性。结果:Ion Torrent PGM第二代测序检测JAK2V617F、CALR和MPL就这3个基因而言,所有样本CALR基因52 bp缺失均未被检测出,其余测序结果与Sanger测序结果完全一致。结论:Lon Torrent PGM测序平台检测MPN患者多个基因突变,方法上具有可行性,能满足临床检测需求。本研究方法在1-2 d内可完成23个基因突变检测,具有灵敏、特异、快速、高通量及低成本的优势。Objective： To investigate the feasibility and relibility of rapidly and accurately acquiring the informations of gene mutations in MPN patients by using self-designed custom MPN mutation-related multipe-PCR primer kit and next generation Ion Torrent PGM sequencing platform. Methods： The bone marrow samples of 10 MPN patients with JAK2V617F and/or CALR＋,Ph-confirmed by sanger sequencing method were collected and were re-detected by using next generation Ion Torrent PGM sequencing method,then the consistence of results of above-mentioned 2 kinds of detection methods was compared. Results： In terms of JAK2V617F,MPL and CALR mutations,the results of Ion Torrent PGM sequencing were complete consistent with results of Sanger sequencing,except 52 bp deletion of CALR gene,which conld not be detected by next generation Ion Torrent PGM sequencing method in all bone marrow samples. Conclusion： The detection of multiple gene mutations in MPN patients by Ion Torrent PGM sequencing platform is feasible and can meet the needs of clinical testing. This method can complete detection of all 23 mutetions within 1-2 days,moreover,possesses advantages of high sensitivity,specificity,rapidity,high throughput and low cost.

关 键 词：骨髓增殖性肿瘤 基因突变 ION Torrent PGM 测序 

分 类 号：R551.3[医药卫生—血液循环系统疾病]