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

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).

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).
- Ratiometric electrochemical biosensor,
- BCR/ABL fusion genes,
- Exonucleases Ⅲ-assisted target recycling,
- Hybrid chain reaction,
- Chronic myeloid leukemia
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沈阳化工大学材料科学与工程学院沈阳 110142