ANALYTICAL CHEMISTRY
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2015
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IF 5.636
Jingpu Zhang,Chao Li,Xiao Zhi,Gabriel Alfranca Ramón,Yanlei Liu,Chunlei Zhang,Fei Pan,Daxiang Cui
MicroRNA (miRNA) biomarkers display great potential for cancer diagnosis and prognosis. The development of rapid and specific methods for miRNA detection has become a hotspot. Herein, hairpin DNA-templated silver nanoclusters (AgNCs/HpDNA) were prepared and integrated into strand-displacement amplification (SDA) as a novel beacon for miRNA detection. The light-up platform was established based on guanine (G)-rich fluorescence enhancement that essentially converted the excitation/emission pair of AgNCs/HpDNAs from a shorter wavelength to a longer wavelength, and then achieved fluorescent enhancement at longer wavelength. On the basis of the validation of the method, the single and duplex detection were conducted in two plasma biomarkers (miR-16-5p and miR-19b-3p) for the diagnosis of gastric cancer. The probe (AgNCs/RED 16(7s)C) utilized for miR-16-5p detection adopted a better conformation with high specificity to recognize single-base mismatches by producing dramatically opposite signals (increase or decrease at 580 nm ex/640 nm em) while the probe (AgNCs/GRE 19b(5s)C) for miR-19b-3p generated dual signals (increase at 490 nm ex/570 nm em and decrease at 430 nm ex/530 nm em) with bright fluorescence in one reaction during the amplification, but unexpectedly was partially digested. This is for the first time to allow the generation of enhanced fluorescent AgNCs and the target recognition integrated into a single process, which offers great opportunity for specific miRNA detection in an easy and rapid way.
研究领域:
生物化学
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分子生物学
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癌症研究
·
纳米技术
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DOI
PubMed
高影响因子
JOURNAL OF ELECTROANALYTICAL CHEMISTRY
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2020
·
IF 3.807
Ziheng Hu,Biying Zhao,Ping Miao,Xialing Hou,Feifei Xing,Yingying Chen,Lingyan Feng
Herein, we present one three-way junction (TWJ) DNA based electrochemical biosensor for sensitive microRNAs detection with locked nucleic acid (LNA) as capture probe and enzyme-free redox cycling signal amplification strategy. LNA owns higher binding affinity and thermal stability compared to normal DNA sequence . In the presence of target microRNA (miRNA-21), novel TWJ structure can be specially triggered with the assistant of methylene blue (MB)-tagged DNA signal probe, thus producing an enhanced electrochemical current. Furthermore, a chemical reductant, tris (2-carboxyethyl) phosphine (TCEP), is adopted to lead a relayed electron transport between solution TCEP and TWJ-linked MB tag and amplified whole signal intensity. Based on the selectivity from target-induced LNA/DNA TWJ structure and enzyme-free MB cycling amplification strategy, the designed electrochemical biosensor exhibits sensitive miRNA-21 detection, with a linear range from 100 aM to 100 pM and a low detection limit of 77 aM. In addition, it can successfully discriminate the differences between the miRNA family members, presenting a promising response to the direct determination of target miRNA-21 in real biological fluids. Our work provides an alternative LNA-based electrochemical biosensor way in the area of miRNA diagnostics and clinical analysis without complicated enzyme/nanomaterials-assisted signal amplification.
研究领域:
生物化学
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分子诊断
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分析化学
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DOI
高影响因子
ADVANCED FUNCTIONAL MATERIALS
·
2024
·
IF 18.5
Yuteng Tang,Yuanhang Xiang,Yu Yang,Yuyuan Zhang,Binqi Wei,Xiaojie Qin,Min Fang,Qiqin Wang,Xinchun Li,Fan Yang
The unmet on-site clinical diagnostics are increasingly demanding portable bioassays, but compromised by sensitivity and throughput. Herein, a nanostructured bubbles-enhanced fluorescence (nBEF) assay is reported by interfacing with smartphone imaging system to enable portable, ultrasensitive, and specific miRNA detection in parallel. In nBEF, the high-curvature bubbles allow physically enhancing optical propagation, chemically enhancing fluorescence, assembling more DNA probes, undergoing faster self-suspension and more efficient self-aggregation fluorescence enhancement relative to their smooth counterparts. By coupling with bubble-driven signal amplification by a plug-and-play DNA concatemer, and with a smartphone imaging system, this nBEF assay achieves a portable and ultrasensitive fluorescent assay of miRNA at the femtomolar level with one-base resolution in a single test, and is capable of monitoring drug-induced miRNA dynamic expression and accurate cancer diagnosis (AUC = 1) through profiling 40 plasma samples after RNA extraction. This assay further integrates an R-value analysis in RGB module with the smartphone to realize automated, on-site miRNA readout. Such a high-performance nBEF assay provides a low-cost, portable, and highly sensitive tool to enable accurate clinical diagnosis.
研究领域:
生物化学
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分子诊断
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医学工程
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临床检验
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DOI
高影响因子
Scientific Reports
·
2025
·Vol. 15
·期 1
·44359
·
IF 3.9
Zhang Yuepeng,Sun Yuhua,Gong Xueshan,Zhao Shuang,Cao Weiwei,Wang Hongbo,Wang Hao,Zhang Xi,Du Changwei,Chen Zhiguo,Deng Lu
Sequence-specific microRNAs (miRNAs) represent promising biomarkers for tumor diagnosis, while current detection methods are often limited by stringent temperature requirements and excessive protease dependency. Herein, we present an isothermal, protein-free cascaded amplification circuit for the sensitive analysis of miRNA-21. This strategy, termed CHA-DNAzyme, integrates a miRNA-activated catalytic hairpin assembly (CHA) reaction with a deoxyribozymes (DNAzyme) component. Specifically, the target miRNA-21 initiates the CHA process via a toehold-mediated strand displacement reaction. This reaction unveils a previously sequestered DNAzyme sequence embedded within a hairpin structure, forming an active double-stranded complex. Subsequently, in the presence of Mg²⁺ cofactors, the catalytically activated DNAzyme drives the continuous cleavage of fluorescent reporter hairpins, releasing a measurable fluorescent signal. Implemented in a one-pot format, the CHA-DNAzyme strategy demonstrates excellent sensitivity (with a limit of detection of 8.70 pM), high specificity (the capability to discern single-base mismatches), short sample-to-answer time (approximately 60 min), and robust performance in analyzing serum samples. This approach holds significant potential to facilitate the advancement of miRNA detection tools and support their application in tumor diagnosis and prognosis assessment.
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PubMed
高影响因子
ANALYTICAL CHEMISTRY
·
2016
·
IF 5.886
Dongqing Kong,Sai Bi,Zonghua Wang,Jianfei Xia,Feifei Zhang
In this work, an in situ growth protocol is introduced to fabricate three-dimensional graphene films (3D GFs) on gold substrates, which are successfully utilized as working electrode for electrochemical detection of nucleic acid (microRNA) and protein (lysozyme) based on a signal-on sensing mechanism. To realize the bridge between the gold substrate and graphene film, a monolayer of 4-aminothiophenol is self-assembled on the substrate, which is then served as connectors for the growth of 3D GFs on the gold substrate by the hydrothermal reduction (HR) technique. Moreover, given the excellent properties, such as enlarged surface area, strong binding strength between 3D GFs and gold substrate, and improved conductivity, the proposed 3D GF-fabricated gold substrate is readily employed to the construction of electrochemical biosensing platforms through introduction of magnetic nanoparticles (MNPs) as probe carriers. On the basis of the strand displacement reaction and specific binding between aptamer and its target, the developed biosensors achieve signal-on detection of microRNA-155 (miR-155) and lysozyme (Lyz) with high sensitivity and selectivity and further successfully applied to human serum assay. Overall, the proposed strategy for in situ growth of 3D GFs provides a powerful tool for a wide range of applications, which is not limited to electrochemical biosensors and can be extended to other areas, such as electrocatalysis and electronic energy-related systems.
研究领域:
材料科学
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分析化学
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生物医学工程
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PubMed
高影响因子
ANALYTICAL CHEMISTRY
·
2019
·
IF 6.35
Huan Gao,Junfang Zhang,Ying Liu,Wenwen Tu,Tianxiang Wei,Zhihui Dai
Development of sensitive detection methods for microRNAs has realistic application value for early clinical analysis and accurate diagnosis. In this study, a triple-helix molecular switch electrochemiluminescence resonance energy transfer (ECL-RET) nanoamplifier was designed to construct an electrochemiluminescence (ECL) biosensor for microRNA determination. The newly synthesized S-doped Lu2O3, which shows a 3 times better ECL performance than Lu2O3, was chosen as the donor in this ECL-RET system. Accordingly, Ag2S quantum dots (QDs) were used as the matched acceptor. They exhibited overlapping spectra and efficient energy transfer between each other. Furthermore, using a triple-helix switch structure with an improved quenching effect for signal amplification and a nano-DNA walker transformational system as a powerful method for target amplification, a supernanoamplifier was achieved. As a consequence, the proposed ECL biosensor for microRNA-141 detection exhibited good analytical performance with a low detection limit (16 aM). The sensor not only led to the development of a novel ECL-RET pair but also provided a strong nanoamplifier for biosensing platform construction, which may offer some new considerations regarding material design, signal amplification, and show promising application value in biomedical and clinical diagnosis.
研究领域:
生物传感技术
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纳米材料科学
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临床诊断
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DOI
PubMed
高影响因子
SPECTROCHIMICA ACTA PART A-MOLECULAR AND BIOMOLECULAR SPECTROSCOPY
·
2022
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IF 4.831
Qingyou Cai,Fanfan Wang,Jingying Ge,Zhiguo Xu,Mei Li,Hui Xu,Hua Wang
Herein, we reported the G-wire-based self-quenched fluorescence probe and its application in ultrasensitive microRNA (miRNA) detection by combining with target-activated isothermal cascade amplification. The terminal-single-fluorescein (FAM)-labeled G-rich oligonucletides self-assembled into G-wire nanostructures (G-wires) with K + and Mg 2+ . Thereafter, the G-wires brought terminal-labeled FAM into close proximity, as a result, the self-quenched signal probe formed. Besides, when there was the target miRNA, target-activated isothermal cascade amplification converted miRNA into the copious trigger DNA. After hybridization between trigger DNA and the self-quenched probe, the G-wires were splited and forced the apart of proximate FAM, and then the self-quenched probe displayed an “on” mechanism. Therefore, the approach gave a limit of detection (LOM) of 0.82 aM to miRNA-21 and could be implemented within a wide linear range of 2 aM to 2 nM. This approach was able to distinguish the single-mismatched miRNA-21, which was selective and sensitive in detecting human spiked serum samples.
研究领域:
分子生物学
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生物化学
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纳米技术
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癌症诊断
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DOI
PubMed
高影响因子
Journal of Pharmaceutical Analysis
·
2025
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IF 8.9
Xinlan Zhu,Ziyan Zhang,Ruiyang Ma,Huachu Chen,Yi Zheng,Su Zeng,Zheyong Li,Sheng Cai
A label-free detection technique for microRNA (miRNA) was developed based on target replacement cycling, polymerization and isomerization cyclic amplification (PICA), and G/Hemin DNAzyme signal output. The designed hairpin probe undergoes cyclic alternation of polymerization and isomerization catalyzed by Bst DNA polymerase, enabling target replacement cycling and continuous self-extension of single-stranded DNA (ssDNA) strands. This process generates abundant G-quadruplex sequences that bind to Hemin, achieving label-free detection of miRNA through catalytic reactions. Its key advantages include a specific primer self-extension mechanism that avoids spurious amplification, one step isothermal design simplifies workflow and enhances system robustness, adapting to diverse detection scenarios, and label-free detection strategy that substantially reduces cost. Therefore, it is poised to become a reliable tool for clinical miRNA analysis, offering potential guidance for the early diagnosis, treatment, and prognosis of tumor biomarkers.
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高影响因子
ANALYST
·
2025
·
IF 3.3
Daqi Chen,Zhezhi Fang,Yachunyue Zhou,Ziyao Zeng,Jiancong Liang,Baian Zhu,Chaozhan Chen
MicroRNAs (miRNAs), as pivotal regulators of gene expression, are critically associated with the progression of cancer, neurodegenerative, and cardiovascular diseases. However, their detection remains challenging due to low sensitivity, background noise, and matrix interferences in complex biological samples, hindering clinical applications such as early tumor screening and liquid biopsy. Herein, we present a cascade-amplified fluorescence biosensor driven by the synergistic integration of exonuclease III (Exo III) and self-locking DNAzyme (SLD). This strategy exploits Exo III-mediated cyclic digestion of target-induced DNA/RNA heteroduplexes to release multiple signal conversion chains, while the SLD’s rigid conformational gate suppresses nonspecific catalysis until unlocked by target-specific molecular keys. The activated DNAzyme then drives secondary amplification through iterative cleavage of fluorophore-labeled reporters. This dual-amplification mechanism achieves label-free, isothermal detection of miRNA-155 with a linear range of 50 fM–10 pM and a limit of detection (LOD) of 4.96 fM. Notably, the sensor demonstrates robust specificity in serum matrices, effectively overcoming homologous miRNA cross-reactivity and complex biological interferences. By merging target recycling with conformation-gated catalysis, this platform offers a cost-effective, high-precision solution for miRNA profiling, holding significant promise for point-of-care diagnostics and liquid biopsy-based early cancer detection.
研究领域:
分子生物学
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肿瘤学
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转化医学
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生物传感
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临床检验
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产品关联
DOI
PubMed
高影响因子
MICROCHIMICA ACTA
·
2018
·Vol. 185
·期 4
·1-8
·
IF 5.705
Yu Xinsheng,Hu Lianzhe,Zhang Feng,Wang Min,Xia Zhining,Wei Weili
A dual-channel ratiometric nanoprobe is described for detection and imaging of microRNA. It was prepared from MoS 2 quantum dots (QDs; with blue emission and excitation/emission peaks at 310/398 nm) which acts as both the gene carrier and as a donor in fluorescence resonance energy transfer (FRET). Molecular beacons containing loops for molecular recognition of microRNA and labeled with carboxyfluorescein (FAM) were covalently attached to the MoS 2 QDs and serve as the FRET acceptor. In the absence of microRNA, the nanoprobe exhibits low FRET efficiency due to the close distance between the FAM tag and the QDs. Hybridization with microRNA enlarges the distance between QD and beacon. This results in an enhancement of the FRET efficiency of the nanoprobe. The ratio of green and blue fluorescence (I 520 /I 398 ) increases linearly in the 5 to 150 nM microRNA concentration range in both aqueous solution and diluted artificial cerebrospinal fluid. The limit of detection (LOD) is as low as 0.38 nM and 0.52 nM, respectively. Other features of this nanoprobe include (a) excellent resistance to nuclease-induced false positive signals and (b) the option to use it for distinguishing different cell lines by in-situ imaging of intracellular microRNAs.
Schematic of a dual-channel photoluminescence nanoprobe for the determination of microRNA-21 (miR-21) by monitoring the microRNA-triggered enhancement of the fluorescence resonance energy transfer (FRET) efficiency between MoS 2 QDs and carboxyfluorescein-labeled molecular beacons.
研究领域:
纳米技术
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生物分子工程
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分析化学
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分子生物学
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产品关联
DOI
PubMed
高影响因子
CHEMICAL ENGINEERING JOURNAL
·
2019
·
IF 8.355
Tongxu Gu,Zhengyan Li,Zhaohui Ren,Xiang Li,Gaorong Han
Simultaneous detection of multiple types of microRNA (miRNA) is of great significance in early cancer diagnosis. In this study, rare-earth-doped CaF 2 upconversion nanocrystals (CaF 2 : RE 3+ , namely Ho 3+ or Tm 3+ and Yb 3+ ) are formed within the mesopores of silica nanoparticles (CaF 2 : RE 3+ @MSN), in an in-situ manner, by capillary effect and thermo-decomposition method. Comparing to current upconversion nanoparticles, the amorphous and mesoporous nature of CaF 2 : RE 3+ @MSN provides abundant active groups and high specific surface area for feasibly immobilizing oligonucleotide probe. The simultaneous and selective detection of multiple miRNAs was achieved based on the photoluminescence spectra of different CaF 2 : RE 3+ @MSN conjugated with DNA probes in the same solution. The results indicated that the proposed approach holds significant potential for multiple miRNA detection and offers an opportunity for cancer early diagnosis.
研究领域:
纳米技术
·
生物医学工程
·
分析化学
·
分子生物学
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产品关联
DOI
高影响因子
Small
·
2020
·Vol. 16
·期 19
·2000460
·
IF 11.459
Manshu Peng,Feifei Sun,Na Na,Jin Ouyang
Nanogap antennas are plasmonic nanostructures with a strong electromagnetic field generated at the gap region of two neighboring particles owing to the coupling of the collective surface plasmon resonance. They have great potential for improving the optical properties of fluorophores. Herein, nanogap antennas are constructed using an aqueous solution-based method to overcome the defects of weak fluorescence and photobleaching associated with traditional organic dyes, and a highly sensitive nanogap antenna-based sensing strategy is presented for the detection of low-abundance nucleic acid biomarkers via a target-triggered strand displacement amplification (SDA) reaction between two DNA hairpins that are tagged to the tips of gold nanorods (Au NRs). In the presence of targets, end-to-end Au NR dimers gradually form, and the fluorophores quenched by the Au NRs exhibit a dramatic fluorescence enhancement due to the plasmon-enhanced fluorescence effect of nanogap antennas. Meanwhile, the SDA reaction results in secondary amplification of fluorescence signals. Combined with single-molecule counting, this method applied in miRNA-21 detection can achieve a low detection limit of 97.2 × 10 −18 m . Moreover, accurate discrimination between different cells through miRNA-21 imaging demonstrates the potential of this method in monitoring the expression level of low-abundance nucleic acid biomarkers.
研究领域:
纳米技术
·
分子生物学
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生物医学工程
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分析化学
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DOI
PubMed
高影响因子
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