Homogeneous electrochemical biosensor for microRNA based on enzyme-driven cascaded signal amplification strategy
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RESEARCH PAPER
Homogeneous electrochemical biosensor for microRNA based on enzyme-driven cascaded signal amplification strategy Yitong Huang 1 & Xiaocui Huang 2 & Huixia Zheng 2 & Cuiying Lin 2 & Zhenyu Lin 2 Received: 19 September 2020 / Revised: 14 October 2020 / Accepted: 23 October 2020 # Springer-Verlag GmbH Germany, part of Springer Nature 2020
Abstract Infectious diseases are a long-standing and severe global public health problem. The rapid diagnosis of infectious diseases is an urgent need to solve this problem. MicroRNA (miRNA) plays an important role in the intervention of some infectious diseases and is expected to become a potential biomarker for the diagnosis and prognosis of infectious diseases. It is of great significance to develop rapid and sensitive methods for detecting miRNA for effective control of infectious diseases. In this study, a simple and highly sensitive homogeneous electrochemical method for microRNAs using enzyme-driven cascaded signal amplification has been developed. In the presence of target miRNA, the reaction system produced plenty of MB-labeled single-nucleotide fragments (MB-MF) containing a few negative charges, which can diffuse to the negative surface of the ITO electrode easily, so an obvious electrochemical signal enhancement was obtained. Without the target, MB-HP contains a relatively large amount of negative charges due to the phosphates on the DNA chain, which cannot be digested by the enzyme and cannot diffuse freely to the negatively charged ITO electrode, so only a small signal was detected. The enhanced electrochemical response has a linear relationship with the logarithm of miRNA concentration in the range of 10 fM to 10 nM and the limit of detection as low as 3.0 fM. Furthermore, the proposed strategy showed the capability of discriminating single-base mismatch and performed eligibly in the analysis of miRNA in cell lysates, exhibiting great potential for disease diagnosis and biomedical research. Keywords Homogeneous . Electrochemical biosensor . MicroRNA . Enzyme-driven cascaded signal amplification . Methylene blue
Introduction MicroRNAs(miRNAs),aclassofnoncodingRNAconsisting of18–25nucleotides,playanessentialroleinphysiologicaland pathological processes [1–3]. MiRNAs have received extensive attention by researchers because of their abnormal Published in the topical collection Analytical Chemistry for Infectious Disease Detection and Prevention with guest editors Chaoyong Yang and XiuJun (James) Li. * Cuiying Lin [email protected] * Zhenyu Lin [email protected] 1
Department of Gynecological Oncology, 26 division, Wenzhou Central Hospital, Wenzhou 325000, Zhejiang, China
2
Ministry of Education Key Laboratory for Analytical Science of Food Safety and Biology, Fujian Provincial Key Laboratory of Analysis and Detection for Food Safety, College of Chemistry, Fuzhou University, Fuzhou 350116, Fujian, China
expression in a variety of diseases and have been regarded as valuablebiomarkersthatcanbeusedinthediagnosisandtreatmentofsomecancers[4–6].Inrecentyears
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