An electrochemical biosensor for detection of PML/RARA fusion gene using capture probe covalently immobilized onto poly-calcon carboxylic acid modified glassy carbon electrode.
In this article, the poly-calcon carboxylic acid (poly-CCA) film modified electrode was prepared by cyclic voltammetry (CV). Then, an electrochemical DNA biosensor was developed for detection of PML/RARA fusion gene in acute promyelocytic leukemia (APL) by using 18-mer single-stranded deoxyribonucleic acid as the capture probe. The capture probe was covalently attached through free amines on the DNA bases using 1-ethyl-3-(3-dimethylaminopropyl) carbodiimide (EDC) and N-hydrosulfosuccinimide (NHS) cross-linking reaction on a carboxylate-terminated poly-CCA monolayer modified glassy carbon electrode (GCE). The covalent immobilized capture probe could selectively hybridize with its target DNA to form double-stranded DNA (dsDNA) on GCE surface. The aim of this work is to provide a well-defined recognition interface for the detection of DNA. Differential pulse voltammetry (DPV) was used to monitor the hybridization reaction on the capture probe electrode. The decrease of the peak current of methylene blue (MB), an electroactive indicator, was observed upon hybridization of the probe with the target DNA. The results indicated that in pH 7.0 phosphate buffer solution (PBS), the oxidation peak current was linear with the concentration of complementary strand in the range of 1.0 x 10(-12) to 1.0 x 10(-11)M with a detection limit of 6.7 x 10(-13)M. This new method demonstrates its excellent specificity for single-base mismatch and complementary sequence (dsDNA) after hybridization, and it would be proposed to use in real sample.
Department of Pharmaceutical Analysis, Fujian Medical University, 88 Jiaotong Road, Fuzhou 350004, China.
Na
N
Wei
Jinghua
J
Chen
Jing
J
Zhang
Kun
K
Wang
Xiongwei
X
Xu
Jianhua
J
Lin
Guangwen
G
Li
Xinhua
X
Lin
Yuanzhong
Y
Chen
D000818:Animals; D015374:Biosensing Techniques / Q000295:instrumentation / Q000379:methods* / Q000592:standards; D015342:DNA Probes; D055664:Electrochemical Techniques; D004566:Electrodes; D052138:Genes, Neoplasm; D006801:Humans; D015473:Leukemia, Promyelocytic, Acute / Q000175:diagnosis; D009693:Nucleic Acid Hybridization; D015514:Oncogene Proteins, Fusion / Q000235:genetics*
D015342:DNA Probes; D015514:Oncogene Proteins, Fusion; C093476:promyelocytic leukemia-retinoic acid receptor alpha fusion oncoprotein