Journal of Materials Chemistry B
·
2024
·
IF 6.1
Miao He,Zhiqiang Hou,Feifan Yin,Wenting Cheng,Yang Xiang,Zhongyun Wang
The detection of a single biomarker is prone to false negative or false positive results. Simultaneous analysis of two biomarkers can greatly improve the accuracy of diagnosis. In this work, we designed a new method for coinstantaneous detection of two breast cancer biomarkers miRNA-21 and HER2 using the properties of duplex-specific nuclease (DSN). Cy5-labeled DNA1 and FAM-labeled DNA2 are used as signal probes to distinguish the two signals. When the sample contains the targets HER2 and miRNA-21, HER2 binds to the HER2 aptamer on the double-stranded DNA2, while miRNA-21 binds to the complementary DNA1. Then, DSN enzyme is added to cut the DNA probes adsorbed on the HER2 aptamer and miRNA-21, releasing the fluorescent groups, which can be readsorbed to the empty sites, thus repeating the cutting of the probes and producing an exponential signal amplification with two distinct fluorescent signals. The detection limits of miRNA-21 and HER2 are 1.12 pM and 0.36 ng mL−1, respectively, with linear ranges of 5 pM to 50 pM and 1 ng mL−1 to 15 ng mL−1. The method was validated in real biological samples, providing a new approach for synchronous analysis of important markers in breast cancer.
研究领域:
生物化学
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分子生物学
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医学诊断
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DOI
PubMed
高影响因子
Molecular Therapy-Nucleic Acids
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2020
·
IF 7.032
Wenjie Mao,Chong Hu,Haifeng Zheng,Jinrong Xie,Xiaorui Shi,Yarong Du,Fu Wang
MicroRNAs (miRNAs) are emerging as vital biomarkers since their abnormal expression is associated with various disease types including cancer. Therefore, it is essential to develop a sensitive and specific platform to monitor the dynamic expression of miRNAs for early clinical diagnosis and treatment. In this study, we designed a functionalized polydopamine (PDA)-based theranostic nanoprobe for efficient detection of miRNA-21 and in vivo synergistic cancer therapy. PDA was modified with polyethylene glycol (PEG) and the obtained PDA-PEG nanoparticles showed good stability in different solutions. PDA-PEG nanoparticles were loaded with fluorescein isothiocyanate (FITC)-labeled hairpin DNA (hpDNA) and an anticancer drug doxorubicin (DOX). In the absence of miRNA-21, PDA effectively quenched the fluorescence of FITC-labeled hpDNA. The presence of miRNA-21 specifically recognized hpDNA and induced the dissociation of hpDNA from PDA-PEG and subsequently recovered the fluorescence signals. Upon cellular uptake of these nanoprobes, a dose-dependent fluorescence activation and synergetic cytotoxic effect were observed due to the release of DOX and inhibition of miRNA-21 function. Furthermore, PDA-PEG-DOX-hpDNA nanoparticles can afford long-term monitoring of miRNA-21 and combined therapeutic efficacy in the nude mice bearing 4T1 tumors. Our results demonstrate the capability of PDA-PEG-DOX-hpDNA as a theranostic nanoprobe for continuously tracking of miRNAs and synergetic cancer therapy.
研究领域:
生物医学工程
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分子生物学
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肿瘤学
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DOI
PubMed
高影响因子
TALANTA
·
2023
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IF 6.1
Kai Zheng,Jiangfei Pan,Zipei Yu,Changqing Yi,Mei-Jin Li
We have proposed a signal dual-amplification electrochemiluminescence (ECL) strategy based on tris(2,2′-bipyridyl)ruthenium(II) (Ru(bpy) 3 2+ ) as chromophores confined with three-dimensional (3D) zinc oxalate metal−organic frameworks (Ru(bpy) 3 2+ @MOFs) for the detection of miRNA-21. The three-dimensional chromophore connectivity in zinc oxalate MOFs provided a network among Ru(bpy) 3 2+ units, shielding the chromophores from solvent molecules and resulting in high Ru(II) complex emission efficiency. Additionally, we discovered that magnetic beads (MBs) used as carrier for enriched signals contribute to enhanced ECL intensity of the chromophore. To evaluate its clinical application, we applied this method to determine the concentration of miRNA-21 solutions ranging from 1.56 to 100 nM, obtaining a calibration curve of ECL intensity versus logarithm of concentration (logC) of miRNA-21 with a high correlation coefficient. This work demonstrates the construction of a signal amplification strategy ECL biosensor for miRNA using Ru(bpy) 3 2+ @MOF systems and its application in ECL detection for analyte methodology.
研究领域:
生物化学
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分子生物学
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医学诊断
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DOI
PubMed
高影响因子
ANALYTICA CHIMICA ACTA
·
2022
·
IF 6.911
Xin Guo,Tian Tian,Xinchun Deng,Yang Song,Xiaoming Zhou,Erqun Song
Development of rapid and accurate detection of miRNAs in complex samples is of great significance for potential early diagnosis of disease. Herein, we report a magnetic relaxation switching (MRS)-based strategy for direct detection of miRNAs in complex samples via the assistance of signal amplification system of CRISPR/Cas13a which has the ability to specifically recognize target RNA. In the designed strategy, 30 nm-magnetic nanoparticles (MB 30 ) and 1000 nm-magnetic particles (MM 1000 ) linked by single-strand RNA 1 complexes (MB 30 -RNA 1 - MM 1000 ) were employed as signal probe. After the target miRNAs (taking miR-21 as model) recognition by CRISPR/Cas13a system, the resulted trans -cleavage degrades the MB 30 -RNA 1 -MM 1000 , releasing MB 30 which caused transverse relaxation time (T 2 ) signal change. The combination of CRISPR/Cas13a assisted signal amplification and the MRS assay achieved direct detection of miR-21 in the serum sample without extracting within 60min, with a detection limit of 0.22 pM. Moreover, the detection accuracy is confirmed by performing the detection of miR-21 using qRT-PCR. The CRISPR/Cas13a system assisted MRS assay successfully achieved accurate, simple, and rapid detection of miRNAs in complex samples, showing great potential for detection miRNAs in potential clinical applications.
研究领域:
分子生物学
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医学诊断
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生物技术
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DOI
PubMed
高影响因子
ANALYTICA CHIMICA ACTA
·
2025
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IF 6
Qian Kai Lin,Wen Qing Sun,Bo Zhang,Qiao Ling,Yan Qian,Cheng Zhi Huang,Chun Mei Li,Na Wang
Background MicroRNAs (miRNAs) play a pivotal role in various physiological and pathological processes. In particular, miRNA-21 holds significant potential as a novel biomarker for the diagnosis of heart failure. The development of miRNA detection methods is rapidly advancing, with point-of-care testing (POCT) platforms garnering considerable attention. However, traditional methods are often hampered by their reliance on expensive instruments and complex procedures, limiting clinical applicability. Therefore, there is an urgent need to develop a simple, portable, sensitive, and rapid POCT platform for miRNA-21 detection. Results In this work, we proposed a portable POCT platform using a colorimetric biosensor specifically sensitive for miRNA-21. The platform utilized 3,3′,5,5′-tetramethylbenzidine as the signaling molecule, and a Linear G-quadruplex loaded with Blocker Nanostructures (LGBN) generated by the RCA reaction as the probe. Furthermore, changes in primary color channels (R/G/B) of TMB for miRNA-21 detection were analyzed via smartphone-based digital image recognition. Under optimal conditions, the platform showed a linear detection range between 0.01 nM and 1 nM, with limits of detection of 8.3 pM (colorimetric methods) and 9.5 pM (digital image colorimetry). Moreover, the colorimetric biosensor exhibited excellent specificity and resistance to interference, successfully detecting miRNA-21 in serum samples from heart failure patients. Significance This detection method has an accuracy consistent with RT-qPCR results, providing a novel and practical approach with POCT for miRNA-21 detection.
研究领域:
分子生物学
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医学诊断
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生物传感技术
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DOI
PubMed
高影响因子
SENSORS AND ACTUATORS B-CHEMICAL
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2024
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IF 8
Jing Xu,Xinqi Luo,Yujin Li,Yinbing Liu,Jianqi Lu,Xi Lin,Ke-Jing Huang
MicroRNA (miRNA) sensitive detection is significant for cancer diagnosis. Currently, self-powered biosensors in clinical miRNA detection show the advantages of no external power supply, mild reaction conditions, low cost and portability. In this work, the bimetallic sulfide heterojunction hollow nanocubes are prepared as the substrate material, which can significantly improve the electrochemical reaction rate and increase the enzyme load. Meanwhile, we design the independent duplex-chain assisted toehold mediated strand displacement reaction (TSDR) strategy. The two substable double chains trigger a continuous chain displacement reaction, and the current signal amplification is realized based on molecular recognition. The constructed sensor has a linear response range of 0.05–10000 fM and a detection limit (LOD) of 8.3 aM. In addition, we use commercial apps to present signals, fundamentally changing the signal output mode of electrochemical sensors, and facilitating the construction of a portable real-time sensor platform. The self-powered biosensor has the characteristics of good biocompatibility, real-time monitoring and economic efficiency in miRNA detection, which provides a new idea for sensitive detection of miRNA. Moreover, it has far-reaching reference significance for the development of other sensing technologies, which is expected to extend from the medical field to environmental monitoring, food safety and other fields.
研究领域:
生物医学工程
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分子诊断
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纳米材料科学
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DOI
高影响因子
JOURNAL OF PHARMACEUTICAL AND BIOMEDICAL ANALYSIS
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2020
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IF 3.209
Shuyu Mei,Bingru Liu,Xiaoli Xiong,Xu Hun
PtCuCo trimetallic alloys (PtCuCo-TAs) are synthesized by one-step reduction. The chemiluminescence (CL) properties of PtCuCo-TAs are studied systemically. PtCuCo-TAs show good catalyzing for luminol-H 2 O 2 system. A CL platform is developed for the detection of miRNA-21 using PtCuCo-TAs as nanozyme catalyzer. In the CL detection platform, H1 (Hairpin DNA1) is immobilized onto magnetic beads (MBs) firstly. In the presence of miRNA-21, H1 is opened. H2 (Hairpin DNA2) then hybridizes with H1. Meanwhile, a “cleat” in the end of miRNA-21 with a fewer bases complementary is formed to prevent miRNA-21 dissociating from H1. This miRNA-21 hybridizes to another H1. When cpDNA-PtCuCo-TAs which consisted with cDNA (Complementary strand of probe DNA) and pDNA-PtCuCo-TAs (PtCuCo-TAs labeled with probe DNA) are added, the ssDNA region of H1 reacts with the toehold domain of probe DNA and cDNA is released resulting pDNA-PtCuCo-TAs being captured. With this process repeatedly, a lot of pDNA-PtCuCo-TAs are captured onto MBs. After separation and washing, the precipitate and H 2 O 2 are put into the 96-well and luminol solution is injected. The CL signal is produced by PtCuCo-TAs catalyzing luminol-H 2 O 2 system. The amount of miRNA-21 is detected with CL signal. This CL platform performs with limit of detection 0.167 fM and has good selectivity over other RNA.
研究领域:
生物化学
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分析化学
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分子生物学
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PubMed
高影响因子
Analytical Methods
·
2019
·Vol. 11
·期 23
·2960-2968
·
IF 2.378
Le Peng,Jun Zhou,Zhaoheng Liang,Yaping Zhang,Lucia Petti,Tao Jiang,Chenjie Gu,Danting Yang,Pasqule Mormile
Based on surface-enhanced Raman scattering (SERS) technology, Au@Ag@4MBA@5′-NH2-ssDNA probes and a Ag/TiO2@3′-NH2-ssDNA substrate were prepared and constructed into a sandwich structure to develop a high sensitivity bioassay of miRNA-21. The Au@Ag@4MBA@5′-NH2-ssDNA probes were prepared by immobilizing 5′-NH2-ssDNA onto the surfaces of 4MBA-labelled Au@Ag core–shell nanoparticles, and the Ag/TiO2@3′-NH2-ssDNA substrate was prepared by immobilizing 3′-NH2-ssDNA on the surface of Ag/TiO2 nanowires SERS-active substrates. The experimental results showed that the SERS-based sandwich bioassay of miRNA-21 presented a low limit of detection of 0.75 fM and a broad dynamic range from 1.0 fM to 1.0 nM. Also, the test data for the SERS-based sandwich bioassay were not only consistent with that of the real-time fluorescence quantitative polynucleotide chain reaction (RT-qPCR) method but also displayed higher detection sensitivity. It was shown that the SERS-based sandwich bioassay of miRNA-21 has importance for use in potential applications involving diagnosing clinical cancer patients.
研究领域:
生物技术
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分子诊断
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纳米医学
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DOI
高影响因子
MICROCHEMICAL JOURNAL
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2025
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IF 5.1
Rong Wang,Junlong Wang,Xin Zhang,Siyu Liao,Yusi Zhou,Qincong Feng,Youjiang Tan,Yuehua Huang,Yuanyuan Dai,Yuanwen Jiang,Haimei Qin
Breast cancer (BC) is the most common malignancy in women and ranks second among all cancer types worldwide. The escalating demand for early and precise diagnosis has intensified the quest for ultrasensitive and highly specific biosensors. Here we present a self-referencing photoelectrochemical (PEC) strategy based on a nanopipette that enables the direct quantification of the BC biomarker miRNA-21 in single living cells. The nanopipette's ultrafine tip minimizes membrane disruption, while an integrated self-referencing electrode cancels out trans-membrane potential fluctuations, yielding a linear dynamic range of 1–7 nM and a detection limit of 0.328 nM without cell lysis or amplification. The platform markedly enhances the accuracy of single-cell heterogeneity analyses and offers a new paradigm for intracellular trace-nucleic-acid quantification. Moreover, the modular self-referencing architecture creates a methodological foundation for extending nanopipette-based sensors to broader single-cell omics applications.
研究领域:
生物化学
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生物传感器
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癌症研究
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纳米技术
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分析化学
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分子生物学
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DOI
高影响因子
Biosensors-Basel
·
2022
·Vol. 12
·期 9
·724
·
IF 5.743
Yang Qing,Yuxing Yang,Ping Ouyang,Chenxin Fang,Haobin Fang,Yazhen Liao,Haiyu Li,Zhencui Wang,Jie Du
Because microRNAs (miRNAs) are biological indicators for the diagnosis, treatment, and monitoring of tumors, cancers, and other diseases, it is significant to develop a rapid, sensitive, and reliable miRNA detection platform. In this study, based on miRNA-21 detection, DNA-a with a 3′ end overhang and Texas Red fluorophore-labeled 5′ end was designed, which reacts with miRNA-21 and hybridizes with exonuclease III (Exo III), where the part connected to miRNA-21 is hydrolyzed, leaving a-DNA. At the same time, miRNA-21 is released to participate in the following reaction, to achieve cyclic amplification. a-DNA reacts with DNA-b conjugated to gold nanoparticles to achieve fluorescence quenching, with the quenching value denoted asF; additionally, after adding DNA-d and linked streptavidin immunomagnetic beads (SIBs), fluorescence recovery was achieved using DNA-c, with the recovered fluorescence recorded asF0. By comparing the difference in the fluorescence (F0−F) between the two experiments, the amount of DNA-a hydrolyzed to produce a-DNA was established to determine the target miRNA-21 content. Under optimized conditions, by comparing the changes in the fluorescence signal, the developed strategy shows good sensitivity and repeatability, with a detection limit of 18 pM, good discriminative ability and selectivity, and promise for the early diagnosis of breast and intestinal cancers.Keywords:exonuclease III;miRNA-21;AuNPs;fluorescence quenching;fluorescence recovery
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
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纳米技术
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生物医学工程
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DOI
PubMed
高影响因子
MICROCHIMICA ACTA
·
2020
·Vol. 187
·期 8
·1-7
·
IF 6.232
Chen Nandi,Li Junyu,Feng Xianzhen,Yang Yanping,Zhu Li,Chen Xiaomeng,Liu Xuan,Li Yang,Wang Cunchuan,Xia Ligang
A label-free fluorescence method based on self-assembled DNA nanopompom has been developed for miRNA-21 detection. In the presence of miRNA-21, three DNA hairpin probes with split G-quadruplex assemble the DNA nanopompom. Based on the isothermal toehold-mediated DNA strand displacement reaction, the target miRNA can be catalytically recycled and trigger three DNA hairpin probes to self-assemble the DNA nanopompom and release the G-quadruplex. The formation of the G-quadruplex increases the fluorescence emission intensity of thioflavin. For thioflavin-based miRNA-21 detection, the excitation and emission wavelengths are set to 425 nm and 490 nm, respectively. The limit of detection for miRNA-21 is 0.8 pM according to F / F 0 = 0.0031 × C miRNA-21 + 1.0382 ( R 2 = 0.9978). This sensing system provides a low-cost, effective, and convenient method for miRNA detection, which holds great potential in biochemical diagnosis and clinical practice.
Label-free and self-assembled fluorescent DNA nanopompom for miRNA detection
研究领域:
生物化学
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纳米技术
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分子生物学
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分析化学
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产品关联
DOI
PubMed
高影响因子
MICROCHEMICAL JOURNAL
·
2025
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IF 5.1
Weiying Liu,Xiaolu Sui,Yunpeng Xu,Tingfei Xie,Yanzi Zhang,Aisha Zhang,Zhenyu Lin,Jihong Chen
MicroRNA-21 (miRNA-21) can serve as an early biomarker for acute kidney injury (AKI), it is important to develop sensitive and selective detection methods for miRNA-21 detection. In this study, an electrochemiluminescence (ECL) biosensor for miRNA-21which synergistically integrates T7 RNA polymerase-driven transcriptional amplification with CRISPR/Cas13a-mediated collateral cleavage and magnetic separation strategy had been designed. T7 RNA polymerase-driven transcriptional amplification has the characters of low requirements for conditions and high amplification efficiency. A cascaded signal amplification strategy which coupled enzymatic target recycling with CRISPR/Cas13a-mediated exponential signal amplification was proposed to achieve high detection sensitivity. And T7 promoter specificity and CRISPR/Cas13a sequence targeting had been adopted to ensure high specificity of the developed system. Furthermore, magnetic bead separation had been adopted to reduce the background signal. The parameters which may affected the performance of the biosensor had been optimized. Under optimized conditions, the ECL response of the system has a linear relationship with the logarithmic value of miRNA-21 concentration in the range of 1.0 fM to 1.0 nM, with a calculated limit of detection(LOD) of 0.33 fM (S/N = 3). The proposed methodology has been adopted to detect the target in clinical urine specimens with satisfied results.
研究领域:
生物化学
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分子生物学
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分析化学
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生物医学工程
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临床诊断
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产品关联
DOI
高影响因子
危险品化学品经营许可证（带存储）