MICROCHIMICA ACTA
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2020
·Vol. 187
·期 5
·1-9
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IF 6.232
Cao Chen,Liu Jinquan,Tang Shuangyang,Dai Zhongran,Xiao Fubing,Rang Weiqing,Liu Ling,Chen Tuo,Yuan Yali,Li Le
A superior electrochemical biosensor was designed for the determination of UO 2 2+ in aqueous solution by integration of DNAzyme and DNA-modified gold nanoparticle (DNA-AuNP) network structure. Key features of this method include UO 2 2+ inducing the cleavage of the DNAzyme and signal amplification of DNA-AuNP network structure. In this electrochemical method, the DNA-AuNP network structure can be effectively modified on the surface of gold electrode and then employed as an ideal signal amplification unit to generate amplified electrochemical response by inserting a large amount of electrochemically active indicator methylene blue (MB). In the presence of UO 2 2+ , the specific sites on DNA-AuNP network structure can be cleaved by UO 2 2+ , releasing the DNA-AuNP network structure with detectable reduction of electrochemical response intensity. The electrochemical response intensity is related to the concentration of UO 2 2+ . The logarithm of electrochemical response intensity and UO 2 2+ concentration showed a wide linear range of 10~100 pM, and the detection limit reached 8.1 pM (S/ N = 3). This method is successfully used for determination of UO 2 2+ in water samples.
Fabricated DNAzyme network structure for enhanced electrical signal. Numerical experiments show that the current signal decreases as the concentration of UO 2 2+ increases. It can be seen that the biosensors could be used to detect UO 2 2+ in aqueous solution effectively.
研究领域:
生物化学
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环境科学
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分析化学
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PubMed
高影响因子
TALANTA
·
2024
·
IF 5.6
Ruili Wang,Ruiying Peng,Liran Song,Jishan Li
Herein, we present a colorimetric sensing strategy for the identification and quantification of tumor-associated miRNAs based on dual DNAzyme amplification. In this sensing ensemble, the substrate portion of the Pb 2+ -dependent 8−17 DNAzyme combines with the G-quadruplex portion to form a hairpin substrate strand. The two split 8−17 DNAzyme strands are partially complementary to the substrate strand and serve as a recognition unit for binding the target miRNA. In the presence of the target miRNA, the activated DNAzyme cleaves the substrate strand, releasing the G-quadruplex. This G-quadruplex binds to hemin to form a G-quadruplex/hemin complex with horseradish peroxidase (HRP)-like properties, which catalyzes the oxidation of ABTS 2− by H 2 O 2 . This oxidation reaction produces a colorimetric signal output, enabling the detection of the target miRNA. Under the optimal reaction conditions explored in this study, the constructed sensing ensembles tailored for each of the specific target miRNAs successfully identified and quantified the four target miRNAs—miR-122, miR-21, miR-335, and miR-155—in both buffer solutions and cell extracts. This colorimetric sensing strategy offers significant advantages in terms of simplicity, cost, and versatility and holds great potential for wide application in biomedical research and clinical diagnostics.
研究领域:
生物化学
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分子诊断学
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分析化学
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PubMed
高影响因子
INTERNATIONAL JOURNAL OF ENVIRONMENTAL ANALYTICAL CHEMISTRY
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2020
·
IF 1.431
Wei Wu,Chundi Yu,Junhua Chen,Qingli Yang
This study aimed to develop a fluorescence assay for Cu2+ monitoring with cascade signal amplification using split Mg2+-dependent DNAzyme subunits as the sensing system and fluorescence as the signal reporter. A subunit of DNAzyme can be formed through a Cu2+-catalysed click reaction between azide- and alkyne-modified short sequences immobilized on magnetic beads, eliminating the washing steps of conventional assays. The immobilized subunit recruits the assisting oligonucleotides to form an autocatalytic DNAzyme as the signal amplifier and a caged fluorescent molecule as the signal reporter. This assay is ultrasensitive, enabling the detection of trace levels of Cu2+ as low as 2 nM, which enhances the detection sensitivity by approximately 100-fold. In addition, the whole reaction process does not require a long manipulation time or a complicated procedure, making the system simple and cost-effective. This method is robust and can be applied to the determination of spiked Cu2+ in water samples with satisfactory recovery. The combination of click chemistry with fluorescent DNAzyme for a simple on-site detection assay can be used for the rapid and sensitive screening of Cu2+.
研究领域:
生物化学
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分析化学
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高影响因子
MICROCHEMICAL JOURNAL
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2021
·
IF 4.821
Qiao Xu,Ke Liu,Jingjing Jin,Xinfeng Zhang
The assay of biomarker proteins is of critical importance for disease diagnosis. In this work, we have designed a cascaded strand displacement reaction system for detection of low abundant biomarker protein in serum samples. The proposed binding-induced strand displacement conversion system, first converts a protein recognition event into the output of the DNA (cDNA) catalyst, and then the output cDNA catalytically load the reporter DNAzyme onto magnetic beads based on catalyzed hairpin assembly (CHA) reaction. The high density of the hairpin DNA on a magnetic bead surface results in an over 20-fold improvement in the reaction efficiency of CHA, compared with conventional CHA. In addition, the magnetic beads are able to efficiently separate the serum sample matrix that could remarkably affect the DNAzyme-catalyzed chemiluminescent (CL) reaction. This type of magnetic biosensor allows for the detection of 0.5 pM target protein in serum samples, whereas when using homogenous CHA biosensors, the CL signal for 2 nM PDGF-BB is seriously inhibited by the serum matrix. Since the sensing system integrates the protein recognition, signaling, signal amplification and matrix separation functions, it is promising for the assay of proteins in biological samples.
研究领域:
生物化学
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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
TALANTA
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2022
·
IF 6.556
Yujie Yu,Min Qing,Yidan Ma,Yan Zhang,Shuting Zheng,Chao Yu,Junlin He
Lead ion (Pb 2+ ) is a major non-degradable environmental pollutant and poses threat to human health. DNAzymes have proved to be a promising tool for the detection of extracellular metal ions . Owing to cellular uptake, enzymatic degradation and reliable signal output, using DNAzymes to quantitative intracellular sensing of Pb 2+ still presents challenges. In this study, we report a core-shell nanoworms-DNAzyme nanoprobe combing hairpin-like DNAzyme (H DNAzyme) with polydopamine-coated Au cores plasmonic “nanoworms” ( [email protected] NWs) for imaging of Pb 2+ in living cells. PDA with good biocompatibility, high near-infrared (NIR) fluorescence quenching efficiency, and provides a larger surface for further functionalization of biomolecules. H DNAzyme can loaded onto [email protected] NWs via coordination interaction and π–π stacking, resulting in significant quenching of their fluorescence by PDA shells and Au cores. In the presence of target Pb 2+ , the H DNAzyme are activated and release Cy5 fluorophore-labeled short strands from [email protected] NWs. Therefore, an increased fluorescence signal is observed and allows imaging of Pb 2+ in living cells. Moreover, the highly selective of H DNAzyme make the proposed method effectively discriminate against other nonspecific molecules. The [email protected] NWs-H DNAzyme nanoprobe shows good biocompatibility and obviously decreased background fluorescence, which can monitor Pb 2+ levels and biodistribution. The developed nanoprobe holds great potential for further extend to bioimaging of other metal ions.
研究领域:
生物化学
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环境科学
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纳米技术
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分子生物学
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DOI
高影响因子
TALANTA
·
2025
·
IF 6.1
Wenchao Ran,Chi Xu,Xiaomei Pan,Xiaojun Huang,Chong Wang,Xianggui Chen,Jie Tang,Imran Mahmood Khan,Yukun Huang
Ofloxacin, a synthetic antimicrobial drug, exhibits significant therapeutic potential in combating various bacterial infections. The exploration of ofloxacin's antimicrobial properties and the unique catalytic activity of G-triplex(G3) DNAzymes demonstrate the continuous efforts to develop cutting-edge biosensors for bacterial detection. G3 outperforms G4 due to a short, easily generated and dissociated sequence, which allows G3 multimeric sequences to fold more accurately than G4 ones of equivalent length, resulting in higher activity for the formed G3/hemin. This study constructed a novel colorimetric DNAzyme biosensor based on G3/hemin for ofloxacin in food. Aptamer-functionalized Fe 3 O 4 magnetic nanoparticles acted as magnetic separation and detection probes, while G3/hemin served as DNAzyme. Under the optimum condition, the presence of ofloxacin initiated the catalytic oxidation to yield a colored product to quantify the ofloxacin concentration with the linear range of 50–300 nmol/L. Under the optimized conditions, a limit of detection of 15.25 nmol/L was obtained by directly detecting the fluorescent signals, which shows obvious superiority over the previously reported methods. This method offers the advantages of simplicity, rapid detection, and high recovery rates in standardization experiments involving actual samples such as beef jerky and ham sausage, paving the way for its widespread implementation in the fields of biomedical diagnostics, environmental monitoring, and food safety.
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高影响因子
TALANTA
·
2024
·
IF 5.6
Yuting Xu,Lili Li,Yumeng Yang,Chengzhi Huang,Hongyan Zou
Developing a simple, reliable, and sensitive hepatitis C virus (HCV) genetic sensing platform is of great significance for diagnosing diseases and selecting appropriate antiviral treatments. Herein, a tandem nucleic acid amplification strategy for sensitive detection of HCV genotype 1b (HCV-1b) was developed by stringing the catalytic hairpin assembly (CHA) and the triggered DNAzyme amplifier. The hairpin reactants were initiated by the target to produce lots of triggering double-stranded DNA sequences which can efficiently activate the subsequent blocked DNAzyme. Thereby, the continuous cleavage of substrate was realized, resulting in the fluorescence signal amplification. The DNA-based isothermal CHA-DNAzyme (CDz) sensing platform was successfully applied for sensitive detection of HCV-1b with the limit of detection (84 pM) and showed good selectivity. Moreover, the practical detection of target DNA in the complex biologic matrix indicated that the developing strategy had good potential for early HCV infection diagnosis.
研究领域:
分子生物学
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病毒学
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诊断学
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PubMed
高影响因子
TALANTA
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2024
·
IF 5.6
Wenqing Shao,Xiaoting Liu,Yi Zhou
Methotrexate (MTX) is a widely used antimetabolite drug, mainly used in the treatment of a variety of cancer. Given the low therapeutic index and significant individual variability of MTX, it was critical to perform therapeutic drug monitoring (TDM) to minimize the side effects. Here, we designed a rapid and sensitive fluorescence/colorimetric assay for the detection of MTX in diluted human serum. After the aptamer binds specifically to MTX, the Walking strand cleaves the Hairpin strand and releases a large amount of fluorescent signal, and the color of the gold nanoparticles changes after the addition of sodium chloride. The change of color could be visualized by eyes to achieve point-of-care TDM of MTX. The quantitative detection of MTX concentration is carried out through the collection of fluorescent signals or absorptions. The fluorescence method can detect MTX in the range of 0.05–1000 μM with the detection limit of 0.0243 μM, meanwhile, the colorimetric method can detect MTX in the range of 0.01–100 μM with the detection limit of 0.0097 μM. The magnetic separation-assisted DNAzyme walker-based nanosensor exhibited good sensitivity, selectivity and stability for detecting MTX in serum and achieve point-of-care methods of TDM of MTX and the nanocarriers has demonstrated significant potential for clinical application.
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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纳米技术
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DOI
PubMed
高影响因子
RSC Advances
·
2019
·Vol. 9
·期 27
·15196-15204
·
IF 3.049
Yun Hong,Dongsen Mao,Rui Wu,Zhe Gao,Tingting Meng,Rongrong Wang,Lin Liu,Jing Miao
DNAzymes have the potential to suppress gene expression through sequence-specific mRNA cleavage and can therefore play an important role in various gene therapies. Hepatitis B virus (HBV) is still one of the most serious liver infections in people around the world and is difficult to treat. We previously designed a 10-23 DNAzyme called DrzBS, which targets HBV S gene expression, but this enzyme depends on exogenous delivery, and so its application has been limited. To overcome this limitation, we have now developed a chitosan-based nanocarrier (chitosan-g-stearic acid, CSO–SA) for intracellular delivery of DrzBS, then compared the inhibition effect of our CSO–SA/DrzBS complex to a common transfection reagent, Lipofectamine™ 2000/DrzBS, on hepatitis B surface antigen expression. The synthesized CSO–SA assembles into micelles in an aqueous solution and exhibits excellent cytoplasmic targeting, and could protect DrzBS from degradation by ribonuclease. CSO–SA/DrzBS showed a higher inhibition rate (IR) than Lipofectamine™ 2000/DrzBS. Moreover, at the same DrzBS concentration (1.2 μmol L−1), the maximum IR of CSO–SA/DrzBS micelles was 2.4-fold that of the Lipofectamine™ 2000/DrzBS complex, and held on for 96 hours. Compared with Lipofectamine™ 2000/DrzBS, CSO–SA/DrzBS achieved a higher HBV inhibition effect. This study demonstrates that CSO–SA micelles can serve as a potential vector for DrzBS and that CSO–SA/DrzBS micelles are a promising application for anti-HBV gene therapy.
研究领域:
医学
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分子生物学
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基因治疗
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高影响因子
BIOSENSORS & BIOELECTRONICS
·
2015
·
IF 6.409
Hongyan Zhang,Ling Lin,Xiaoxue Zeng,Yajuan Ruan,Yongning Wu,Minggui Lin,Ye He,FengFu Fu
We herein developed a novel biosensor for the visual detection of trace uranyl ion ( UO 2 2 + ) in aqueous environment with high sensitivity and specificity by using DNAzyme-functionalized magnetic beads (MBs) for UO 2 2 + recognition and gold nano-particles (AuNPs)-based enzymatic catalysis oxidation of TMB (3,3′,5,5′-tetramethylbenzidine sulfate) for signal generation. The utilization of MBs facilitates the magnetic separation and collection of sensing system from complex sample solution, which leads to more convenient experimental operation and more strong resistibility of the biosensor to the matrix of sample, and the utilization of AuNPs-based enzymatic catalysis amplification greatly improved the sensitivity of the biosensor. Compared with the previous DNAzyme-based UO 2 2 + sensors, the proposed biosensor has outstanding advantages such as relative high sensitivity and specificity, operation convenience, low cost and more strong resistibility to the matrix of sample. It can be used to detect as low as 0.02 ppb (74 pM) of UO 2 2 + in aqueous environment by only naked-eye observation and 1.89 ppt (7.0 pM) of UO 2 2 + by UV–visible spectrophotometer with a recovery of 93–99% and a RSD≤5.0% ( n =6) within 3 h. Especially, the visual detection limit of 0.02 ppb (74 pM) is much lower than the maximum allowable level of UO 2 2 + (130 nM) in the drinking water defined by the U.S. Environmental Protection Agency (EPA), indicating that our method meets the requirement of rapid and on-site detection of UO 2 2 + in the aqueous environment by only naked-eye observation.
研究领域:
环境科学
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生物传感
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纳米技术
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PubMed
高影响因子
ANALYTICAL CHEMISTRY
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2020
·
IF 6.785
Xiangdan Meng,Kai Zhang,Fan Yang,Wenhao Dai,Huiting Lu,Haifeng Dong,Xueji Zhang
MicroRNAs (miRNAs) are involved in the essential progresses of many diseases and have emerged as therapeutic and diagnostic biomarkers. The combination of miRNA aberrant expression and tumor microenvironment (TME) features holds great potential for precise tumor imaging diagnosis but has been minimally explored. Herein, we rationally design a DNA@Cu-MOF nanosystem containing copper metal–organic frameworks (Cu-MOF) and a DNAzyme-assisted signal amplification procedure for deregulated miRNA-related hypoxic tumor diagnosis. The nanoprobes comprising a signal strand block Cu-specific DNAzyme precursor and a substrate strand are assembled on the surface of the hypoxia-responsive Cu-MOF. Under TME characterized by hypoxia, the DNA@Cu-MOF nanosystem disassociates and accomplishes the release of abundant Cu2+, DNAzyme precursor, and substrate strand. Target aberrant miRNA displaces the signal strand to recover one fluorescence signal for detection. Importantly, it activates the Cu-specific DNAzyme amplification, which produces miRNA aberrant expression-dependent fluorescence signal for hypoxic tumor diagnosis. Both in vitro and in vivo experiments validate its good performance for tumor cell diagnosis. The hypoxia-driven and miRNA-binding-induced self-powered and temporal-spatial fluorescence imaging nanosystem not only provides a great tool for aberrant miRNA-related hypoxic tumor diagnosis but also is readily applied for the control and modulation of biological functions.
研究领域:
生物医学工程
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分子生物学
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纳米医学
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DOI
高影响因子
ANALYTICA CHIMICA ACTA
·
2023
·
IF 6.2
Yang Lu,Yiyue Yu,Yeru Wang,Wenbo Zhou,Ziyi Cheng,Liandong Yu,Shaojiang Zheng,Rongke Gao
Background Precise and specific miRNA detection plays a vital role in exploring development mechanisms of cancer disease, thereby it can significantly improve relevant prevention and treatment strategies. Results In this work, a surface-enhanced Raman spectroscopy (SERS)-based microfluidic chip has been devised with a microcone array SERS substrate (MCASS) for the miR-141 detection. This substrate excels in unique SERS activity and large surface area for DNA oligonucleotide modification. As the presence of miR-141, the DNAzyme walker induced cleavage reaction took place on the finely designed and prepared dual DNA conjugated SERS nanoprobes. The SERS nanoprobes can anchor on MCASS by the DNA hybridization that achieved an impressive detection limit in the femtomolar level. Significance With this integrated SERS-based microfluidic chip, we provided a miRNA detection strategy using DNAzyme walker amplification technology. It is believed that this strategy could be a powerful tool for miRNA detection and related cancer screening test .
研究领域:
生物医学工程
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分子生物学
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纳米技术
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DOI
PubMed
高影响因子
危险品化学品经营许可证（带存储）