新型比率型电化学生物传感器的构建及对BCR/ABL融合基因的检测

陈美君 刘雅兰 王颖 戈芷琪 张修华 王升富 何汉平

引用本文: 陈美君, 刘雅兰, 王颖, 戈芷琪, 张修华, 王升富, 何汉平. 新型比率型电化学生物传感器的构建及对BCR/ABL融合基因的检测[J]. 分析化学, 2022, 50(5): 692-700. doi: 10.19756/j.issn.0253-3820.210477 shu
Citation:  CHEN Mei-Jun,  LIU Ya-Lan,  WANG Ying,  GE Zhi-Qi,  ZHANG Xiu-Hua,  WANG Sheng-Fu,  HE Han-Ping. A Novel Ratiometric Electrochemical Biosensor for Detection of BCR/ABL Fusion Gene[J]. Chinese Journal of Analytical Chemistry, 2022, 50(5): 692-700. doi: 10.19756/j.issn.0253-3820.210477 shu

新型比率型电化学生物传感器的构建及对BCR/ABL融合基因的检测

    通讯作者: 何汉平,E-mail:hehanping@hubu.edu.cn
  • 基金项目:

    省部共建生物催化与酶工程国家重点实验室开放基金项目(No.SKLBEE2020016)和国家自然科学基金项目(Nos.52171177,21575035)资助。

摘要: 采用普鲁士蓝(PB)和硫堇(Thi)两种电化学信号物质,同时引入核酸外切酶Ⅲ(Exo Ⅲ)辅助目标循环和杂交链式反应(HCR)两种信号放大策略,构建了一种超灵敏比率型电化学生物传感器,用于BCR/ABL融合基因的检测。首先,通过恒电位沉积法在玻碳电极表面形成一层普鲁士蓝(PB)薄膜,在其上固定捕获DNA。然后,将发夹DNA (H1)与目标DNA杂交配对,在Exo Ⅲ作用下,目标DNA被释放,可进入下一次循环,同时释放出发夹DNA的部分单链片段(Trigger-DNA),此单链可被PB膜和辅助DNA修饰电极捕获,继而在此修饰电极表面进行HCR反应,形成典型的Y形线性DNA串联体。此时,PB的电化学信号降低;另一方面,形成的串联体与Thi通过静电作用结合后,呈现高的Thi电化学响应。Thi和PB的电流响应信号比值(IT/IP)的对数与目标DNA的浓度对数在1 pmol/L~100 nmol/L范围内具有良好的线性关系,线性方程为lg (IT/IP)=1.094×lgC+0.156,检出限为0.19 pmol/L。本方法对BCR/ABL融合基因具有较高的选择性。此比率型电化学生物传感器为慢性粒细胞白血病(CML)的诊断和预后提供了一种简单可行的策略。

English


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  • 收稿日期:  2021-04-30
  • 修回日期:  2022-01-28
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