ANALYTICAL CHEMISTRY
·
2025
·
IF 6.7
Wenting Cheng,Han Pan,Jinhua Chen,Miao He,Zhongyun Wang,Yang Xiang
Exosomal miRNAs, particularly miRNA-21, are promising cancer biomarkers. Current enzyme-dependent detection methods face challenges, such as environmental limitations and high costs. In contrast, enzyme-independent sensors are highly desirable for on-site, miniaturized, and cost-effective miRNA detection. To address these limitations, we developed a nonenzymatic electrochemical sensor featuring a triple-signal amplification system for ultrasensitive detection of miRNA-21. This sensor utilizes cascade toehold-mediated strand displacement reactions to activate molecular machines triggered by target miRNA, generating biotinylated-and-thiol-modified double-stranded DNA for stable immobilization on a gold electrode. Preprepared biotinylated tetrahedron DNA (TDNA)-mediated hybridization chain reaction probes are then linked to the electrode via streptavidin–biotin binding. This amplification process allows for significant DNA duplex immobilization, with electroactive [Ru(NH3)6]3+ (RuHex) adsorbed onto them, producing a robust electrochemical signal. This approach enables accurate detection of miRNA-21 at concentrations as low as 0.43 fM, with a linear range from 1 fM to 1 nM. Clinical testing demonstrates its potential for cancer diagnostics.
研究领域:
生物化学
·
医学诊断
·
分子生物学
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产品关联
DOI
PubMed
高影响因子
Analytical Methods
·
2019
·Vol. 11
·期 26
·3286-3293
·
IF 2.378
Zhiqiang Jiao,Lei Zhao,Chu Tang,Hongyan Shi,Fu Wang,Bo Hu
Recently, 3D printing technology has been widely applied in microfluidics due to its capacity of three-dimensional construction, rapid and automatic fabrication, and less time consumption. However, a droplet-based PCR in 3D-printed microfluidic chips is rarely reported. In this study, a 3D-printed microfluidic chip was designed and fabricated for droplet-based PCR detection of miRNA-21. In this chip, the capacity of monodisperse droplet generation was demonstrated. Moreover, the temperature of the chip was uniform and sufficiently stable, meeting the requirement for the PCR process. In addition, the working conditions of the droplet-based PCR process in the 3D-printed chip were explored. Finally, the droplet-based PCR detection of miRNA-21 levels in the MCF-10A cells and MDA-MB-231 cells was successfully achieved using the 3D-printed chip. We expect that the enormous potential of 3D printing technology in the research area of microfluidics will be explored in the future.
研究领域:
微流控技术
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分子生物学
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生物医学工程
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DOI
高影响因子
SENSORS AND ACTUATORS B-CHEMICAL
·
2026
·
IF 8.3
Xuan Weng,Guanbo Wang,Longyan Chen,Hai Jiang
This study presents the development of a microfluidic thread-based electrochemical nanobiosensor for the ultrasensitive detection of the lung cancer biomarker microRNA-21 (miR-21). The biosensor was constructed on nylon thread electrodes modified with poly(3,4-ethylenedioxythiophene):polystyrene sulfonate (PEDOT:PSS)–MXene–gold nanoparticles (AuNPs) (PMA) nanocomposites. Capture probes (CPs) were immobilized on the thread electrode surface via Au-S bonds. A robust signal amplification was achieved through horseradish peroxidase (HRP) catalysis, activated upon the formation of a sandwich hybridization structure between the target miR-21 and biotin-labeled signal probes (SPs). The electrochemical properties of the nanobiosensors were characterized using cyclic voltammetry (CV) and electrochemical impedance spectroscopy (EIS). Under optimized conditions, the biosensor exhibited a wide linear detection range from 1 fM to 1 nM, with an ultralow detection limit of 0.15 fM. It also demonstrated high specificity against mismatched sequences, good reproducibility, and acceptable stability. When applied to spiked human serum samples, the recovery rates ranged from 96.4% to 104.3%, confirming its potential for clinical sample analysis. The thread-based three-electrode system offers advantages over conventional screen-printed and glassy carbon electrodes, including adjustable electrode spacing and layout flexibility, low cost and a simple fabrication process. The proposed method provides a promising technological platform for developing sensitive, stable, and point-of-care (POC) diagnostic tools for miR-21 and other disease-related biomarkers.
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DOI
高影响因子
ELECTROCHIMICA ACTA
·
2026
·
IF 5.8
Yunna Li,Zhidan Tian,Liang Zhou,Jinming Kong,Xueji Zhang
Target-triggered CHA generates azide-bearing duplexes on the GCE surface. • CuAAC links CHA-derived DNA assemblies with ATRP initiator sites. • Py-mediated photo-ATRP forms electroactive poly(FMMA) for signal amplification. • The platform shows a 100 aM–100 pM range and a blank-based LOD of 60 aM. • Matrix-matched calibration supports preliminary analysis in 10% diluted NHS. The electrochemical conversion of nucleic acid recognition events into amplified current signals remains important for developing sensitive miRNA biosensors. Herein, we propose a target-programmed and light-triggered interfacial signal-amplification strategy for miRNA-21 detection by coupling catalytic hairpin assembly (CHA), CuAAC click chemistry, and pyrene-mediated photo-induced atom transfer radical polymerization (photo-ATRP). In this design, miRNA-21 first triggers CHA on the H1-modified glassy carbon electrode surface, generating surface-bound H1–H2 duplexes carrying terminal azide groups. These azide-bearing duplexes are subsequently coupled with the alkyne-containing ATRP initiator PBIB through CuAAC, thereby converting the target-triggered DNA assembly product into interfacial polymerization-related sites. Under UV irradiation, pyrene activates the photo-ATRP of ferrocenylmethyl methacrylate (FMMA), leading to the in situ formation of electroactive poly(FMMA) and an enhanced square-wave voltammetry response. The stepwise sensing mechanism was supported by agarose gel electrophoresis, FTIR, component-deletion SWV experiments, EIS/CV characterization, and interfacial morphology analysis. Under optimized conditions, the biosensor showed a linear response toward miRNA-21 from 100 aM to 100 pM, with a blank-based limit of detection of 60 aM. The method also exhibited acceptable selectivity, reproducibility, short-term storage stability, and preliminary recovery performance in 10% diluted normal human serum spiked samples with matrix-matched calibration. This work demonstrates a proof-of-concept electrochemical amplification strategy that links target-catalytic DNA assembly with light-controlled interfacial polymer growth for sensitive miRNA detection. Download: Download high-res image (210KB) Download: Download full-size image
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DOI
高影响因子
CHINESE JOURNAL OF ANALYTICAL CHEMISTRY
·
2021
·
IF 1.134
Qiu-Mei FENG,Li QIN,Peng ZHANG,Dan LI,Ming-Kai LIU,Po WANG
Building a precise, sensitive, and selective sensing platform is highly significant for microRNA (miRNA) detection. Herein, an innovative ratiometric electrochemical biosensor sensitized with carbon nanoparticles-functionalized SnS 2 nanoflowers ( [email protected] 2 NFs) and enzyme-assisted target recycling was explored for analysis of miRNA-21. In this strategy, the application of hierarchical [email protected] 2 NFs structure as electrode material not only enhanced the surface loading capability for subsequent reactions but also improved the interfacial electron transfer. After introducing enzyme-assisted target recycling, single-target miRNA-21 could initiate the cleavage of multiple capture DNA strands by duplex-specific nuclease (DSN), leading to numerous rest single-strand DNAs on the electrode. Two probes, methylene blue-labeled probe 1 (probe 1-MB) and ferrocene-labeled probe 2 (probe 2-Fc), could selectively and competitively hybridize with the capture DNA and the rest single-strand DNA, depending on the completeness of DNA hybridization. By measuring the peak current intensity ratio of Fc and MB, miRNA-21 was quantified in a wide concentration range from 1 fmol/L to 10 pmol/L, with a detection limit of 300 amol/L. Moreover, this sensing system showed good analytical performance in the presence of interfering sequences and complex matrices, illuminating excellent selectivity and practical application potential of the hierarchical [email protected] 2 NF sensing interface.
研究领域:
生物化学
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纳米医学
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分子诊断
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产品关联
DOI
高影响因子
COMPOSITES COMMUNICATIONS
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2024
·
IF 6.5
Minghui Xu,Xiaoru Yao,Yichen Wang,Haihan Liu,Wenjia Yang,Pingping Zhang
In this paper, core-sheath nanofiber membrane (miRNA-21@DCH) co-loaded with microRNA-21 and doxycycline (DCH) was prepared by coaxial electrospinning technology for the treatment of diabetic wounds. DCH could inhibit the growth of Gram-negative and Gram-positive bacteria that easily cause infection at the wound site, thus promoting the epidermal layer thickening and dermal connective tissue hyperplasia. The cumulative release rate of DCH was 89 % after 72 h. In vitro antibacterial experiments at 24 h showed that the bacteriostatic zone diameter of miRNA-21@DCH nanofiber membrane was about 3.82 cm for Escherichia coli and 4.54 cm for Staphylococcus aureus , indicating that miRNA-21@DCH nanofiber membrane had good antibacterial effect. The microRNA-21 could promote angiogenesis and collagen deposition at the wound site, providing a favorable growth environment for tissue cell proliferation. The synergistic effect of DCH and microRNA-21 confers miRNA-21@DCH core-sheath nanofiber membrane the advantage of rapid healing of diabetic wounds. After 16 days treatment with miRNA-21@DCH nanofiber membrane, the wound basically healed and the wound surface was smooth. In addition, in vitro cytotoxicity assays showed that miRNA-21@DCH nanofiber membrane had good biocompatibility. The prepared miRNA-21@DCH nanofiber membrane provides a new and effective strategy for the synergistic treatment of diabetic wounds.
研究领域:
生物材料学
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药理学
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再生医学
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产品关联
DOI
高影响因子
BIOSENSORS & BIOELECTRONICS
·
2023
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IF 12.6
Xia Qiu,Jingyi Dong,Qingshan Dai,Mimi Huang,Yongxin Li
Enzyme-free hybridization chain reaction (HCR) technology is often used as a signal amplification tool for the detection of different targets. In this study, an ultrasensitive and label-free method for detecting miRNA-21 was developed using the nanopore ionic current rectification (ICR) technology coupled with HCR technology. The probe oligonucleotide (DNA1) was combined with the gold-coated nanopore through the Au–S bond to form a DNA1-functionalized gold-coated nanopore (DNA1-Au-coated nanopore). Since miRNA-21 is partially complementary to DNA1, it can be selectively recognized by DNA1-functionalized gold-coated nanopores. The target (miRNA-21) can induce the opening of hairpin DNA and HCR reaction after the introduction of hairpin DNA H1 and H2. The concentration of miRNA-21 will affect the combination of H1 and H2 on the inner wall of the nanopore, and its surface charge will change with the internal modification, thereby changing the ion current rectification ratio . Under the condition that the concentration of H1, H2 and HCR reaction time are constant, the change of ICR ratio is linearly correlated with the logarithm of miRNA-21 concentration within a certain range, which shows that the sensing strategy we designed can achieve target miRNA-21 detection. This ultrasensitive miRNA holds great promise in the field of cancer diagnosis.
研究领域:
生物化学
·
分子生物学
·
纳米技术
·
医学诊断
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产品关联
DOI
PubMed
高影响因子
Nanoscale
·
2020
·Vol. 12
·期 47
·24100-24106
·
IF 6.895
Xia Li,Fang Yang,Chunfang Gan,Ruo Yuan,Yun Xiang
The simultaneous imaging of the dynamic expression variations of regulatory RNAs in cells, which remains a major challenge, has important applications in precise disease diagnosis, treatment and prognosis. Here, we describe the establishment of a biodegradable ZnO nanoparticle (NP)-assisted asymmetric amplification approach for the simultaneous imaging of microRNA-21 (miRNA-21) and programmed cell death 4 (PDCD4) mRNA at distinct expression levels in live cells. The DNA signal probe complexes are immobilized on the ZnO NPs and readily delivered into the target cancer cells via the endocytosis pathway. The acidic microenvironment in cancer cells leads to the dissolution of the ZnO NPs to release Zn2+ ions and the intracellular miRNA-21 activates the Zn2+-dependent DNAzyme to cleave the substrate signal probes with the assistance of the Zn2+ cofactor to show green fluorescence for imaging miRNA-21. Meanwhile, the PDCD4 mRNA can displace the other quenched signal probes to generate red fluorescence. Importantly, the PDCD4 mRNA sequences can be recycled and reused by using the DNAzyme-cleaved sequences as the fuel strands through two strand displacement reactions to yield amplified red fluorescence for detecting low levels of PDCD4 mRNA. Moreover, our approach can be used to evaluate the varied expression levels of miRNA-21 and PDCD4 mRNA responsive to different drugs in cells, reflecting its usefulness for precise cancer diagnosis and prognosis upon anticancer drug treatment.
研究领域:
分子生物学
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癌症研究
·
纳米医学
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DOI
PubMed
高影响因子
ANALYTICAL AND BIOANALYTICAL CHEMISTRY
·
2020
·Vol. 412
·期 23
·5779-5787
·
IF 3.637
Dai Panpan,Ke Jiajun,Xie Chenggen,Wei Liyun,Zhang Ying,He Yong,Chen Lijuan,Jin Juncheng
A sensitive electrochemiluminescence (ECL) assay for microRNAs (miRNAs) based on a semiconductor nanomaterial sensitized with noble-metal Au nanoparticles (NPs) is successfully developed. TiO 2 nanotubes (NTs) were equipped with Au NPs to obtain an enhanced ECL emitter. Then, an ECL assay for miRNA-21 was fabricated, which was based on the use of probe 2 DNA–functionalized Pt/PAMAM nanocomposites (NCs) assembled on the surface of Au/TiO 2 NT conjugate via DNA hybridization between probe 1 DNA and capture DNA. The Pt/PAMAM NCs act as an ECL quencher of Au/TiO 2 NTs via resonance energy transfer. After the binding of target miRNA-21 and the capture DNA, the Pt/PAMAM NCs were released and the ECL signal was recovered. An “off–on” ECL assay was achieved with a linear response from 0.01 to 10,000 pM. Finally, this method has been validated to be sensitive and specific for miRNAs in human serum samples. The ECL enhancement strategy opens a new way for fabricating various sensitive biosensors.
A sensitive “off–on” electrochemiluminescence analysis method was developed, which combined Au NP–enhanced ECL emission of TiO 2 nanotubes and an efficient energy-transfer system between Au/TiO 2 nanotubes and Pt/PAMAM nanocomposites
研究领域:
生物医学工程
·
分析化学
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产品关联
DOI
PubMed
高影响因子
ANALYTICAL CHEMISTRY
·
2024
·
IF 6.7
Shiwei Xu,Guofeng Wang,Yueyue Feng,Juanjuan Zheng,Liying Huang,Jiahao Liu,Yisha Jiang,Yajun Wang,Nannan Liu
MicroRNAs (miRNAs) are endogenous and noncoding single-stranded RNA molecules with a length of approximately 18–25 nucleotides, which play an undeniable role in early cancer screening. Therefore, it is very important to develop an ultrasensitive and highly specific method for detecting miRNAs. Here, we present a bottom-up assembly approach for modifying glass microtubes with silica nanowires (SiNWs) and develop a label-free sensing platform for miRNA-21 detection. The three-dimensional (3D) networks formed by SiNWs make them abundant and highly accessible sites for binding with peptide nucleic acid (PNA). As a receptor, PNA has no phosphate groups and exhibits an overall electrically neutral state, resulting in a relatively small repulsion between PNA and RNA, which can improve the hybridization efficiency. The SiNWs-filled glass microtube (SiNWs@GMT) sensor enables ultrasensitive, label-free detection of miRNA-21 with a detection limit as low as 1 aM at a detection range of 1 aM–100 nM. Noteworthy, the sensor can still detect miRNA-21 in the range of 102–108 fM in complex solutions containing 1000-fold homologous interference of miRNAs. The high anti-interference performance of the sensor enables it to specifically recognize target miRNA-21 in the presence of other miRNAs and distinguish 1-, 3-mismatch nucleotide sequences. Significantly, the sensor platform is able to detect miRNA-21 in the lysate of breast cancer cell lines (e.g., MCF-7 cells and MDA-MB-231 cells), indicating that it has good potential in the screening of early breast cancers.
研究领域:
生物医学工程
·
分子生物学
·
纳米技术
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产品关联
DOI
PubMed
高影响因子
Scientific Reports
·
2016
·Vol. 6
·期 1
·1-9
·
IF 5.228
Miao Xiangmin,Ning Xue,Li Zongbing,Cheng Zhiyuan
Positively charged gold nanoparticles (+)AuNPs can adsorb onto the negatively charged surface of single-stranded DNA (ssDNA) or double-stranded DNA (dsDNA). Herein, long-range dsDNA polymers could form based on the hybridization chain reaction (HCR) of two hairpin probes (H 1 and H 2 ) by using miRNA-21 as an initiator. (+)AuNPs could adsorb onto the negatively charged surface of such long-range dsDNA polymers based on the electrostatic adsorption, which directly resulted in the precipitation of (+)AuNPs and the decrease of (+)AuNPs absorption spectra. Under optimal conditions, miRNA-21 detection could be realized in the range of 20 pM-10 nM with a detection limit of 6.8 pM. In addition, (+)AuNPs used here are much more stable than commonly used negatively charged gold nanoparticles ((−)AuNPs) in mixed solution that contained salt, protein or other metal ions. Importantly, the assay could realize the detection of miRNA in human serum samples.
研究领域:
纳米技术
·
分析化学
·
生物传感器
·
分子诊断
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产品关联
DOI
PubMed
高影响因子
Analytical Methods
·
2016
·Vol. 8
·期 48
·8492-8497
·
IF 1.915
Wenjun Ouyang,ZhenHua Liu,GuiFeng Zhang,Zheng Chen,Longhua Guo,Zhenyu Lin,Bin Qiu,Guonan Chen
MicroRNAs (miRNAs) have attracted enormous interest due to their involvement in complex regulatory functions and for being closely linked to some cancers. Thus, qualitative and quantitative detection of miRNAs has become an urgent necessity. In this study, we report an enzyme-free fluorescent biosensor for miRNA-21 detection based on MnO2 nanosheets and catalytic hairpin assembly amplification. Results showed that the detection limit for miRNA-21 was 0.33 nM (S/N = 3) and the linearity range was from 1 to 50 nM. Compared with traditional methods, such strategy is convenient, rapid, and low-cost and it can have a promising potential for miRNA detection in cancer.
研究领域:
生物化学
·
分子生物学
·
癌症研究
·
纳米技术
✓ 已验证
DOI
高影响因子
危险品化学品经营许可证（带存储）