摘要
表皮生长因子受体(EGFR)的基因是肿瘤检测的生物标志物,基于微滴式数字PCR技术建立EGFR突变基因的检测方法并进行了方法学比较。利用分子生物学方法自主构建了定性定量快速检测人EGFR基因突变的微滴式数字PCR检测方法,应用于80例临床血浆/组织样本的检测;同时与荧光定量PCR技术、第二代DNA测序技术进行了方法学对比。基于微滴式数字PCR建立的检测方法灵敏度达到0.1%,准确度及特异性符合率均为100%。使用不同方法检测了80例临床血浆/组织样本,本方法与荧光定量PCR技术检测一致率为96.25%,与第二代DNA测序技术检测一致率为100%。建立的微滴式数字PCR检测EGFR突变的方法具有高灵敏度、高特异性、快速检测的优势,能够满足临床应用需求,为临床癌症早期诊断及指导肿瘤靶向用药提供理论及技术基础。
The epidermal growth factor receptor(EGFR)gene was a biomarker for tumor detection.Based on micro-droplet digital PCR,a method for detecting EGFR mutant genes were established and compared.A molecular microbiological method was used to construct a micro-droplet digital PCR detection method for qualitative and quantitative rapid detection of human EGFR gene mutations.It was applied to the detection of 80 clinical plasma/tissue samples.Simultaneously with real-time PCR,second-generation DNA Sequencing techniques were compared in methodologies.The sensitivity of the detection method based on micro-drop digital PCR was 0.1%;and the accuracy and specificity were 100%.80 clinical plasma/tissue samples were used for different methods.The coincidence rate of this method and real-time PCR was 96.25%;and the agreement rate with the second-generation DNA sequencing technology was 100%.The established method of microdrop digital PCR detection of EGFR mutation has the advantages of high sensitivity,high specificity and rapid detection,which can meet the needs of clinical application and provide theoretical and technical basis for early diagnosis and guidance of tumor targeting drugs.
作者
孙婷婷
石乐民
徐任
孙彦波
于源华
SUN Ting-ting;SHI Le-min;XU Ren;SUN Yan-bo;YU Yuan-hua(School of Life Science and Technology,Changchun University of Science and Technology,Changchun 130022;Shanghai Ruiang Gene Technology Co.,Ltd.Fengxian District,Shanghai 201400)
出处
《长春理工大学学报（自然科学版）》
2020年第6期135-142,共8页
Journal of Changchun University of Science and Technology(Natural Science Edition)
基金
吉林省科技厅项目(20190304114YY,182466YY010154577,182473TC010551396)。
作者简介
孙婷婷(1994-),女,硕士研究生,E-mail:632097785@qq.com;通讯作者:于源华(1963-),女,博士,教授,E-mail:yuyuanhua8888@126.com。
