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

1. [1]
  FIALKOW P J, GARTLER S M, YOSHIDA A. Proc. Natl. Acad. Sci. U.S.A., 1967, 58(4):1468-1471.FIALKOW P J, GARTLER S M, YOSHIDA A. Proc. Natl. Acad. Sci. U.S.A., 1967, 58(4):1468-1471.
2. [2]
  KANTARJIAN H M, DEISSEROTH A, KURZROCK R, ESTROV Z, TALPAZ M. Blood, 1993, 82(3):691-703.KANTARJIAN H M, DEISSEROTH A, KURZROCK R, ESTROV Z, TALPAZ M. Blood, 1993, 82(3):691-703.
3. [3]
  DALEY G Q, VANETTEN R A, BALTIMORE D. Science, 1990, 247(4944):824-830.DALEY G Q, VANETTEN R A, BALTIMORE D. Science, 1990, 247(4944):824-830.
4. [4]
  LIN X H, WU P, CHEN W, ZHANG Y F, XIA X H. Talanta, 2007, 72(2):468-471.LIN X H, WU P, CHEN W, ZHANG Y F, XIA X H. Talanta, 2007, 72(2):468-471.
5. [5]
  KANTARJIAN H M, KEATING M J, TALPAZ M, WALTERS R S, SIMTH T L, CORK A, MCCREDIE K B, FREIREICH E J. Am. J. Med., 1987, 83(3):445-454.KANTARJIAN H M, KEATING M J, TALPAZ M, WALTERS R S, SIMTH T L, CORK A, MCCREDIE K B, FREIREICH E J. Am. J. Med., 1987, 83(3):445-454.
6. [6]
  HAMBLIN T J. Leuk. Res., 2002, 26(6):607-609.HAMBLIN T J. Leuk. Res., 2002, 26(6):607-609.
7. [7]
  LANDSTROM A P, TEFFERI A. Leuk. Lymphoma, 2006, 47(3):397-402.LANDSTROM A P, TEFFERI A. Leuk. Lymphoma, 2006, 47(3):397-402.
8. [8]
  MVLLER C, HENNIG E, FRANKE C, KRAHL R, LEIBLEIN S, NIEDERWIESER D, DEININGER M W. Cancer Genet. Cytogenet., 2002, 136(2):149-150.MVLLER C, HENNIG E, FRANKE C, KRAHL R, LEIBLEIN S, NIEDERWIESER D, DEININGER M W. Cancer Genet. Cytogenet., 2002, 136(2):149-150.
9. [9]
  CORRENTE F, BELLESI S, METAFUNI E, PUGGIONI P L, MARIETTI S, CIMINELLO A M, ZA T, SORA F, FIANCHI L, SICA S, STEFANO V D, CHIUSOLO P. Cytometry, Part B, 2018, 94(3):468-476.CORRENTE F, BELLESI S, METAFUNI E, PUGGIONI P L, MARIETTI S, CIMINELLO A M, ZA T, SORA F, FIANCHI L, SICA S, STEFANO V D, CHIUSOLO P. Cytometry, Part B, 2018, 94(3):468-476.
10. [10]
  VROTSOS E, GORGAN M, DIGIUSEPPE J A. Cytometry, Part B, 2018, 92(4):275-278.VROTSOS E, GORGAN M, DIGIUSEPPE J A. Cytometry, Part B, 2018, 92(4):275-278.
11. [11]
  WU J L, HUANG Y, BIAN X, LI D, CHENG Q, DING S. Opt. Commun., 2016, 377:24-32.WU J L, HUANG Y, BIAN X, LI D, CHENG Q, DING S. Opt. Commun., 2016, 377:24-32.
12. [12]
  XU Y, BIAN X, SANG Y, LI Y, LI D, CHENG W, YIN Y, JU H, DING S. Sci. Rep., 2016, 6:32370.XU Y, BIAN X, SANG Y, LI Y, LI D, CHENG W, YIN Y, JU H, DING S. Sci. Rep., 2016, 6:32370.
13. [13]
  GULATI P, KAUR P, RAJAM M V, SRIVASTAVA T, ALI M A, MISHRA P, ISLAM S S. Sens. Actuators, B, 2018, 270:45-55.GULATI P, KAUR P, RAJAM M V, SRIVASTAVA T, ALI M A, MISHRA P, ISLAM S S. Sens. Actuators, B, 2018, 270:45-55.
14. [14]
  GHOLIVAND M B, AKBARI A. Biosens. Bioelectron., 2019, 129:182-188.GHOLIVAND M B, AKBARI A. Biosens. Bioelectron., 2019, 129:182-188.
15. [15]
  CIFTCI S, CANOVAS R, NEUMANN F, PAULRAJ T, NILSSON M, CRESPO G A, MADABOOSI N. Biosens. Bioelectron., 2020, 151:112002.CIFTCI S, CANOVAS R, NEUMANN F, PAULRAJ T, NILSSON M, CRESPO G A, MADABOOSI N. Biosens. Bioelectron., 2020, 151:112002.
16. [16]
  WANG Cun, LIU Fan-Xin, HUI Jun-Min, ZOU Xiao-Chuan, DENG Ming-Chuan, ZHENG Tian-Ping, LIU Lin. Chin. J. Anal. Chem., 2020, 48(9):1185-1192. 王存, 刘蕃鑫, 惠俊敏, 邹晓川, 邓明川, 郑天平, 刘林. 分析化学, 2020, 48(9):1185-1192.
17. [17]
  GE L, WANG W, LI F. Anal. Chem., 2017, 89(21):11560-11567.GE L, WANG W, LI F. Anal. Chem., 2017, 89(21):11560-11567.
18. [18]
  GAI P P, GU C C, LI H Y, SUN X Z, LI F. Anal. Chem., 2017, 89(22):12293-12298.GAI P P, GU C C, LI H Y, SUN X Z, LI F. Anal. Chem., 2017, 89(22):12293-12298.
19. [19]
  ZHU L P, ZHANG M Q, YE J, YAN M X, ZHU Q J, HUANG J S, YANG X R. Anal. Chem., 2020, 92(12):8614-8622.ZHU L P, ZHANG M Q, YE J, YAN M X, ZHU Q J, HUANG J S, YANG X R. Anal. Chem., 2020, 92(12):8614-8622.
20. [20]
  ZHAO L, SUN R J, HE P, ZHANG X R. Anal. Chem., 2019, 91(22):14773-14779.ZHAO L, SUN R J, HE P, ZHANG X R. Anal. Chem., 2019, 91(22):14773-14779.
21. [21]
  ZHANG W, LIU C, HAN K, WEI X O, XU Y W, ZHOU X B, ZHANG H, CHEN Z Y. Biosens. Bioelectron., 2020, 154:112091.ZHANG W, LIU C, HAN K, WEI X O, XU Y W, ZHOU X B, ZHANG H, CHEN Z Y. Biosens. Bioelectron., 2020, 154:112091.
22. [22]
  GE L, WANG W X, SUN X M, HOU T, LI F. Anal. Chem., 2016, 88(4):2212-2219.GE L, WANG W X, SUN X M, HOU T, LI F. Anal. Chem., 2016, 88(4):2212-2219.
23. [23]
  MIAO J C, DU K, LI X, XU X T, DONG X, FANG J L, CAO W, WEI Q. Biosens. Bioelectron., 2021, 171:112713.MIAO J C, DU K, LI X, XU X T, DONG X, FANG J L, CAO W, WEI Q. Biosens. Bioelectron., 2021, 171:112713.
24. [24]
  CUI L, LU M F, LI Y, TANG B, ZHANG C Y. Biosens. Bioelectron., 2018, 102:87-93.CUI L, LU M F, LI Y, TANG B, ZHANG C Y. Biosens. Bioelectron., 2018, 102:87-93.
25. [25]
  LI Y R, CHANG Y Y, MA J, WU Z Y, YUAN R, CHAI Y Q. Anal. Chem., 2019, 91(9):6127-6133.LI Y R, CHANG Y Y, MA J, WU Z Y, YUAN R, CHAI Y Q. Anal. Chem., 2019, 91(9):6127-6133.
26. [26]
  HU R, ZHANG X, CHI K N, YANG T, YANG Y H. ACS Appl. Mater. Interfaces, 2020, 12(27):30770-30778.HU R, ZHANG X, CHI K N, YANG T, YANG Y H. ACS Appl. Mater. Interfaces, 2020, 12(27):30770-30778.
27. [27]
  LI J, LIU Y L, ZHU X Q, CHANG G, HE H P, ZHANG X H, WANG S F. ACS Appl. Mater. Interfaces, 2017, 9(50):44231-44240.LI J, LIU Y L, ZHU X Q, CHANG G, HE H P, ZHANG X H, WANG S F. ACS Appl. Mater. Interfaces, 2017, 9(50):44231-44240.
28. [28]
  LIU Y L, GE Z Q, CHEN M J, HE H P, ZHANG X H, WANG S F. Biosens. Bioelectron., 2019, 142:111537.LIU Y L, GE Z Q, CHEN M J, HE H P, ZHANG X H, WANG S F. Biosens. Bioelectron., 2019, 142:111537.
29. [29]
  JIN H, GUI R J, YU J B, LV W, WANG Z H. Biosens. Bioelectron., 2017, 91:523-537.JIN H, GUI R J, YU J B, LV W, WANG Z H. Biosens. Bioelectron., 2017, 91:523-537.
30. [30]
  ZHAO C Q, JIN H, GUI R J, WANG Z H. Sens. Actuators, B, 2017, 242:71-78.ZHAO C Q, JIN H, GUI R J, WANG Z H. Sens. Actuators, B, 2017, 242:71-78.
31. [31]
  YU J B, JIN H, GUI R J, WANG Z H, GE F. Talanta, 2017, 162:435-439.YU J B, JIN H, GUI R J, WANG Z H, GE F. Talanta, 2017, 162:435-439.
32. [32]
  JIANG J, WU H Y, SU Y, LIANG Y, SHU B W, ZHANG C S. Anal. Chem., 2020, 92(11):7708-7716.JIANG J, WU H Y, SU Y, LIANG Y, SHU B W, ZHANG C S. Anal. Chem., 2020, 92(11):7708-7716.
33. [33]
  YUAN Y L, HU T, ZHONG X, ZHU M H, CHAI Y Q, YUAN R. ACS Appl. Mater. Interfaces, 2020, 12(20):22624-22629.YUAN Y L, HU T, ZHONG X, ZHU M H, CHAI Y Q, YUAN R. ACS Appl. Mater. Interfaces, 2020, 12(20):22624-22629.
34. [34]
  GUO Q Q, YU Y Q, ZHANG H, CAI C X, SHEN Q M. Anal. Chem., 2020, 92(7):5302-5310.GUO Q Q, YU Y Q, ZHANG H, CAI C X, SHEN Q M. Anal. Chem., 2020, 92(7):5302-5310.
35. [35]
  WANG J R, XIA C, YANG L, LI Y F, LI C M, HUANG C Z. Anal. Chem., 2020, 92(5):4046-4052.WANG J R, XIA C, YANG L, LI Y F, LI C M, HUANG C Z. Anal. Chem., 2020, 92(5):4046-4052.
36. [36]
  BI S, YUE S Z, ZHANG S S. Chem. Soc. Rev., 2017, 46(14):4281-4298.BI S, YUE S Z, ZHANG S S. Chem. Soc. Rev., 2017, 46(14):4281-4298.
37. [37]
  FENG C R, ZHANG C, GUO J X, LI G P, YE B X, ZOU L N. Anal. Chim. Acta, 2021, 1158:338413.FENG C R, ZHANG C, GUO J X, LI G P, YE B X, ZOU L N. Anal. Chim. Acta, 2021, 1158:338413.

扫一扫看文章

计量

PDF下载量: 1
文章访问数: 36
HTML全文浏览量: 3

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

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？

计量

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？

计量

文章相关

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