Sensitive detection of microRNA using a label-free copper nanoparticle system with polymerase-based signal amplification
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RESEARCH PAPER
Sensitive detection of microRNA using a label-free copper nanoparticle system with polymerase-based signal amplification Jiangnan Lu 1 & Shuping Liang 1 & Li Tan 1 & Kun Hu 1 & Shulin Zhao 1 & Jianniao Tian 1 Received: 19 October 2019 / Revised: 21 July 2020 / Accepted: 27 July 2020 # Springer-Verlag GmbH Germany, part of Springer Nature 2020
Abstract The abnormal expression of microRNAs (miRNAs) has been reported in many diseases, so it is of great interest to develop simple and accurate methods for the detection and analysis of miRNA expression. We have developed a novel biosensor to detect miRNAs. This method is based on a polymeric double-stranded DNA (dsDNA) copper nanoparticle (CuNP) template that is synthesised by a polymerase. When Cu2+ and ascorbic acid are added to the system, the dsDNA template (which is rich in A-T bases) promotes the formation of CuNPs, resulting in high fluorescence intensity. This system provides sensitive analysis of miRNA expression with a limit of detection down to 17.8 pmol/L, due to significant changes in the fluorescence signal of the system before and after the addition of the target. The linear range between F0-F and concentration of miR-122 is 80.0 pmol/L to 4.50 nmol/L, and the recovery rate in spiked HepG2 cell lysates is 93.33–102.53%. This method expands the applications of fluorescent DNA-CuNPs in the field of biosensor analysis, and can be used to detect and analyse any miRNA marker by changing the target recognition sequence. Keywords MicroRNA . Enzymatic polymerisation . Copper nanoparticles . Ascorbic acid
Introduction MicroRNAs (miRNAs) are small, endogenous, non-coding, single-stranded deoxyribonucleotide molecules that are ubiquitous in eukaryotes [1, 2]. They possess great potential as a biological marker for the detection of diseases, especially cancer [3–5]. For example, elevated miRNA-122 expression is closely associated with hepatitis, liver cancer, lung cancer, and other common diseases [3, 6–8]. As the incidence of cancer increases, there is an urgent need for advanced detection methods for miRNA biomarkers in vivo. Because of the low abundance and instability of miRNA in biological samples, constructing a sensitive, selective, and quantitative analysis method is challenging. Electronic supplementary material The online version of this article (https://doi.org/10.1007/s00216-020-02850-y) contains supplementary material, which is available to authorized users. * Jianniao Tian [email protected] 1
Key Laboratory for the Chemistry and Molecular Engineering of Medicinal Resources (Ministry of Education of China), School of Chemistry and Pharmaceutical Science of Guangxi Normal University, Guanxi, Guilin 541004, China
In order to address this challenge, researchers have developed a number of nucleic acid signal amplification techniques for the sensitive detection of miRNAs [9–12], such as polymerase chain reaction (PCR) [13, 14], rolling circle amplification (RCA) [15–18], hybridisation chain reaction (HCR) [19–22], strand displacement amplifi
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