一种突变型循环肿瘤DNA荧光检测方法的构建  

Construction of A Fluorescent Detection Method for Mutant Circulating Tumor DNA

作　　者：包京京 李雨鸣 陈志平 夏江浩 田媛媛 于妍妍 李瑾 BAO Jing-jing;LI Yu-ming;CHEN Zhi-ping;XIA Jiang-hao;TIAN Yuan-yuan;YU Yan-yan;LI Jin(School of Medical Technology,Xuzhou Medical University,Jiangsu Xuzhou 221004;The First School of Clinical Medicine,Xuzhou Medical University,Jiangsu Xuzhou 221004;School of Nursing,Xuzhou Medical University,Jiangsu Xuzhou 221004;School of Pharmacy,Xuzhou Medical University,Jiangsu Xuzhou 221004,China)

机构地区：[1]徐州医科大学医学技术学院,江苏徐州221004 [2]徐州医科大学第一临床医学院,江苏徐州221004 [3]徐州医科大学护理学院,江苏徐州221004 [4]徐州医科大学药学院,江苏徐州221004

出　　处：《广州化工》2024年第11期53-55,共3页GuangZhou Chemical Industry

基　　金：江苏省高等学校大学生实践创新训练计划项目(No:202110313031Z)。

摘　　要：首先合成二氧化硅包覆Fe_(3)O_(4)磁性纳米粒子(Fe_(3)O_(4)-SiO_(2)),然后连接目标物T790M的识别核酸链(DNA1),制备T790M的捕获探针(Fe_(3)O_(4)-SiO_(2)-DNA1)。进一步加入荧光素FAM标记的DNA2探针。当T790M存在时,DNA2,Fe_(3)O_(4)-SiO_(2)-DNA1与T790M发生DNA杂交。磁性分离后,上清液中DNA2上连接的FAM的荧光强度显著下降,且荧光强度与T790M浓度之间具有良好的线性相关性,线性回归方程为F=-142.56lgC+739.24(R^(2)=0.994),检测限为0.02 nmol/L。本方法为恶性肿瘤的精确诊断提供了一种有效的技术。A capture probe for T790M(Fe_(3)O_(4)-SiO_(2)-DNA1)was firstly prepared by synthesizing silica-coated Fe_(3)O_(4)magnetic nanoparticles(Fe_(3)O_(4)-SiO_(2))and attaching the recognition nucleic acid strand(DNA1)of the target T790M.A fluorescein FAM-labelled DNA2 probe was added to the above probe.When T790M was presented,DNA2 and Fe_(3)O_(4)-SiO_(2)-DNA1 underwent DNA complementary hybridization with T790M.After magnetic separation,the fluorescence intensity of FAM attached on DNA2 in the supernatant was significantly reduced,and there was a good linear correlation between fluorescence intensity and T790M concentration,with a linear regression equation of F=-142.56lgC+739.24(R^(2)=0.994)and a detection limit of 0.02 nmol∙L^(-1).The fluorescence assay constructed in this work provided an effective technique for the accurate diagnosis of malignant tumors.

关 键 词：循环肿瘤DNA(ctDNA) DNA杂交 磁性分离 荧光分析 恶性肿瘤 

分 类 号：O657.3[理学—分析化学] R730.4[理学—化学]