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

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

陈美君 ^2, ,
刘雅兰 ^2, ,
王颖 ^2, ,
戈芷琪 ^2, ,
张修华 ^2, ,
王升富 ^2, ,
何汉平 ^{1,2,
,}

1.
湖北大学省部共建生物催化与酶工程国家重点实验室, 武汉 430062;
2.
湖北大学化学化工学院, 武汉 430062

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（I_T/I_P）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 （I_T/I_P）=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。

关键词:

English

A Novel Ratiometric Electrochemical Biosensor for Detection of BCR/ABL Fusion Gene

1.
State Key Laboratory of Biocatalysis and Enzyme Engineering, Hubei University, Wuhan 430062, China;
2.
College of Chemistry and Chemical Engineering, Hubei University, Wuhan 430062, China

Corresponding author: HE Han-Ping, hehanping@hubu.edu.cn

Received Date: 30 April 2021
Revised Date: 28 January 2022
Fund Project:
Supported by the Open Project Funding of the State Key Laboratory of Biocatalysis and Enzyme Engineering (No.SKLBEE2020016) and the National Natural Science Foundation of China (Nos.52171177, 21575035).

Abstract: An ultrasensitive ratiometric electrochemical biosensor was developed for detection of BCR/ABL fusion gene relied on Prussian blue (PB) and thionine (Thi) as double signals. Exonucleases Ⅲ (Exo Ⅲ)-assisted target recycling and hybrid chain reaction (HCR) signal amplification strategies were applied. Above all, the PB film was formed on the electrode surface by controlled potential electrolysis. Exo Ⅲ-assisted target recycling was operated onto the hairpin DNA, which included the specific recognition fragment of BCR/ABL fusion genes according to target DNA-triggered exonuclease reaction, and release of the hairpin fragment. The released hairpin fragments were captured on PB film-modified electrode for further amplification of HCR signal with Help-DNA, AD1 and AD2, which induced the formation of a typical linear DNA concatamers with Y-shaped structure. The formed concatamers were bound with Thi based on electrostatic interaction for second important electrochemical response. The signal ratio (I_T/I_P) of Thi and PB showed an excellent linearity with BCR/ABL fusion gene (lg(I_T/I_P)=1.094 × lgC+0.1557) in the concentration range of 1 pmol/L-100 nmol/L with a detection limit (S/N=3) of 0.19 pmol/L. The developed ratiometric genosensor provided a simple and feasible strategy for diagnosis and prognosis of chronic myeloid leukemia (CML).

Key words:

Ratiometric electrochemical biosensor
/ BCR/ABL fusion genes
/ Exonucleases Ⅲ-assisted target recycling
/ Hybrid chain reaction
/ Chronic myeloid leukemia

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通讯作者: 陈斌, bchen63@163.com

1.
沈阳化工大学材料科学与工程学院沈阳 110142

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

English

A Novel Ratiometric Electrochemical Biosensor for Detection of BCR/ABL Fusion Gene

Corresponding author: HE Han-Ping, hehanping@hubu.edu.cn

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通讯作者: 陈斌, bchen63@163.com

中国化学会秘书处

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

English

A Novel Ratiometric Electrochemical Biosensor for Detection of BCR/ABL Fusion Gene

Corresponding author: HE Han-Ping, hehanping@hubu.edu.cn

CCS Chemistry：嵌段共聚物自组装成 “三明治”二维有机材料，实现纳米级压力传感功能

CCS Chemistry：颜徐州、鲍哲南： 你的皮肤可能是一片SPMs

CCS Chemistry：工作高效不疲劳，只须让催化剂动起来

CCS Chemistry：静电组装导向聚合- 聚电解质纳米凝胶量化制备新策略

CCS Chemistry：金黄色葡萄球菌醛基脱氢酶是如何识别特异性底物的？

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通讯作者: 陈斌, bchen63@163.com

中国化学会秘书处

CCS Chemistry：颜徐州、鲍哲南：你的皮肤可能是一片SPMs