EGFR、ALK、KIT及KRAS基因Panel体细胞突变高通量测序检测法室内质量控制的建立  

Establishment of Internal Quality Control by High-Throughput Sequencing with the Somatic Mutation Panel Composed of EGFR,ALK,KIT and KRAS Genes

作　　者：吴小延[1] 李丹 杨鑫华 刘小云 龙亚康 王芳[1] 邓玲[1] Wu Xiaoyan;Li Dan;Yang Xinhua;Liu Xiaoyun;Long Yakang;Wang Fang;Deng Ling Sun(Yat-sen University Cancer Center,State Key Laboratory of Oncology in South China,Guangzhou 510060,Guangdong,China;Department of Nuclear Medicine,Cancer Center of Guangzhou Medical University,Guangzhou 510095,Guangdong,China)

机构地区：[1]中山大学肿瘤防治中心,华南肿瘤学国家重点实验室,广州510060 [2]广州医科大学附属肿瘤医院核医学科,广州510095

出　　处：《肿瘤预防与治疗》2020年第10期821-829,共9页Journal of Cancer Control And Treatment

基　　金：广东省基础与应用基础研究基金(编号:2020A1515010314,2017A030310192);中山大学青年教师培育项目(编号:17ykpy84)。

摘　　要：目的:利用由295个靶基因所组成的高通量测序Panel,试建立适用于包括表皮生长因子受体(epidermal growth factor receptor,EGFR)、间变淋巴瘤激酶(anaplastic lymphoma kinase,ALK)、酪氨酸激酶受体(tyrosine kinase receptor,KIT)及大鼠肉瘤病毒癌基因(Kirsten rat sarcoma viral oncogene,KRAS)体细胞突变高通量测序法日常检测的室内质量控制体系。方法:选择EGFR、ALK、KIT和KRAS 4种基因7个突变类型的患者样本,按比例稀释成5.00%、2.50%突变频率的样本作为外部质控品,对其均一性、稳定性及特异性进行评价。结果:均一性评价结果显示质控品均一性良好,稳定性评价结果表明质控品检测结果随时间延长无明显趋势变化,且经变异分析均显示变异系数<10%。结论:临床检测突变样本型模式的阳性混合标本制备定量检测的5.00%和2.50%的两个质控品均一性、稳定性及特异性良好,可有效用于基于探针捕获的包括EGFR、ALK、KIT及KRAS基因Panel体细胞突变高通量测序技术检测流程的室内质量控制,检测结果能反映标本的真实性,有一定的临床实用价值。Objective:To establish an internal quality control system suitable for high-throughput sequencing with a somatic mutation panel composed of 295 target genes for the daily detection of epidermal growth factor receptor(EGFR),anaplastic lymphoma kinase(ALK),Tyrosine kinase receptor(KIT)and Kirsten rat sarcoma viral oncogene(KRAS).Methods:We selected samples of EGFR,ALK,KIT and KRAS genes in 7 mutation types,and diluted the samples with the mutation frequency of 5.00%and 2.50%as the external quality control products,and evaluated their homogeneity,stability and specificity.Results:Quality control products had good homogeneity,stability and specificity,which did not change significantly over time(coefficient of variation:<10%).Conclusion:Quality control products with the mutation frequency of 5.00%and 2.50%have good homogeneity,stability and specificity,which can be effectively used for the internal quality control of high-throughput sequencing technology with the somatic mutation panel composed of EGFR,ALK,KIT and KRAS genes based on probe-based capturing.The results can reflect the authenticity of the specimen and have certain practical value in clinic.

关 键 词：高通量测序技术 室内质量控制 质控品 

分 类 号：R730.5[医药卫生—肿瘤] R319[医药卫生—临床医学]