二代测序技术在儿童T淋巴母细胞淋巴瘤全外显子测序中的应用  

Application of next generation sequencing in the whole exome sequencing for pediatric T lymphoblastic lymphoma

作　　者：姚新原 温贤浩[1] 宪莹[1] 朱进[2] 肖剑文[1,3] YAO Xinyuan;WEN Xianhao;XIAN Yin;ZHU Jin;XIAO Jianwen(Blood tumor center,Children Hospital Affiliated to Chongqing Medical University;Key Laboratory of the Ministry of Education for Researchon Child Developmental Diseases;Chongqing Key Laboratory of Pediatrics,Chongqing 400014,China)

机构地区：[1]重庆医科大学附属儿童医院血液肿瘤中心 [2]儿童发育疾病研究教育部重点实验室 [3]儿科学重庆市重点实验室,重庆400014

出　　处：《分子影像学杂志》2018年第4期534-537,共4页Journal of Molecular Imaging

摘　　要：目的建立从石蜡包埋的儿童T淋巴母细胞淋巴瘤(T-LBL)病理组织中提取基因组DNA,采用二代测序技术行全外显子测序检测的方法并分析其可行性。方法从10例儿童T-LBL石蜡包埋组织中提取基因组DNA,PCR反应扩增后采用二代测序技术行全外显子测序检测并确定致病位点,所获得的致病突变采用Sanger测序法测序确认,比较两种方法的一致性。结果 10例石蜡包埋T-LBL组织标本中均成功提取到基因组DNA,采用二代测序技术开展全外显子测序测序均检测到致病突变;二代测序检测结果经Sanger测序法验证,证实致病突变确实存在,且与Sanger测序结果一致。结论基于二代测序技术的全外显子测序测序可用于检测石蜡包埋的儿童T-LBL病理标本的致病突变。Objective To establish the methods of extract DNA and detect whole exome sequencing （WES） by using nextgeneration sequencing （NGS） technique from paraffin embedded samples for children with T lymphoblastic lymphoma （T-LBL） and to explore its feasibility. Methods Genomic DNA was extracted from 10 cases of paraffin embedded samples withpediatric T-LBL. Genomic DNA samples were amplified by polymerase chain reactions, NGS technique was used for WESdetection and pathogenic mutations were founded. The pathogenic mutations were confirmed by Sanger sequencing,comparison of consistency between NGS technique and Sanger sequencing was evaluated. Results Genomic DNA sampleswere successfully extracted from the 10 T-LBL specimens of paraffin embedded tissues. The pathogenic mutations weredetected by NGS technique and the result was consistent with the Sanger sequencing method. Conclusion NGS technologycan be used for WES and detection of pathogenic mutation for paraffin embedded pediatric T-LBL specimens.

关 键 词：石蜡包埋组织 儿童 T淋巴母细胞淋巴瘤 二代测序技术 全外显子测序 

分 类 号：R733.1[医药卫生—肿瘤]