TALANTA
·
2024
·
IF 5.6
Yingao Yang,Ruipeng Chen,Yifen Guo,Jiaxin Zhang,Shuyue Ren,Huanying Zhou,Zhixian Gao
A two-color fluorescent sensing strategy based on a functionalized tetrahedral DNAzyme nanotweezer (FTDN) was developed to detect ochratoxin A (OTA) utilizing the multifunctional properties of DNA nanotechnology. The FTDN enables rapid OTA detection directly through a Cy5 fluorescent group, modified to respond to the target signal. Additionally, FTDN exhibits DNAzyme cutting activity in the presence of Mg 2 ⁺ ions, enabling it to traverse DNA nanoflower-functionalized magnetic beads. This process results in the continuous cleavage of DNA nanoflowers labeled with numerous FAM fluorescent groups, thereby amplifying the detection signal and enhancing OTA sensitivity. The linear ranges for the Cy5 and FAM signals in response to OTA were 5–1000 ng/mL and 0.05–100 ng/mL, respectively, with corresponding limits of detection (LOD) of 1.59 ng/mL and 0.03 ng/mL. This study demonstrates that dual-color fluorescence via Cy5 and FAM can effectively verify OTA detection in food, significantly reducing false-positive and false-negative rates. The proposed platform offers sensitive and accurate detection of mycotoxins in food and can be adapted for monitoring other trace contaminants by simply altering the aptamer.
研究领域:
分析化学
·
纳米技术
·
食品科学
·
毒理学
✓ 已验证
产品关联
DOI
PubMed
高影响因子
SENSORS AND ACTUATORS B-CHEMICAL
·
2014
·
IF 3.84
Zi-Ming Zhou,Yong Yu,Ming-Zhen Zhang,Jun Chen,Qiong-Qiong Ren,Ji-Tao Song,Bo Liu,Zhi-Ya Ma,Yuan-Di Zhao
It is important to design new ways with high specificity and sensitivity to detect lead (II). Here, ‘8–17’ DNAzyme was used as a Pb 2+ -dependent probe with high sensibility and selectivity based on chemiluminescence (CL). When Pb 2+ was present, the substrate strand was cleaved and the active hemin/G-quadruplex DNAzyme structure was formed in the presence of another sequence which contained two parts, one could be complementary with part of the substrate strand and the other was a G rich subunit. The active hemin/G-quadruplex DNAzyme structure could catalyze luminol oxidation and quantitative analysis of Pb 2+ was carried out according to the CL of luminol. Results showed that CL intensity of luminol at 452 nm had a linear relation with the concentration of Pb 2+ in the range from 5 nM to 1 μM, and the limit of detection was 0.79 nM ( S / N = 3). This method was also used to detect the contents of Pb 2+ in lake water and tobacco and the results (62.4 ± 4.0 nM and 15.2 ± 2.2 nM, respectively) were extremely closed to those of inductively coupled plasma-mass spectrometry (ICP-MS) (57.0 ± 1.4 nM and 14.5 ± 0.91 nM, respectively).
研究领域:
生物化学
·
环境科学
·
分析化学
✓ 已验证
产品关联
DOI
高影响因子
FOOD RESEARCH INTERNATIONAL
·
2022
·
IF 7.425
Li Zou,Manjun Zhang,Mengyan Li,Zhiyou Xiao,Liansheng Ling
This work developed an enzyme-free fluorescent aptasensor for sensitive aflatoxin B1 (AFB1) detection based on a dual signal amplification strategy of hybridization chain reaction (HCR) and Zn 2+ -dependent DNAzyme. In the presence of AFB1, the aptamer specifically binds to the target, releasing the blocking DNA, which can initiate HCR between hairpin probes H1 and H2. With the addition of the substrate strand (Zn-Sub) and enzyme strand (Zn-Enz) of DNAzyme, HCR product can hybridize with Zn-Sub and Zn-Enz to form triplex DNA and Y-shaped structure together, which further activates the DNAzyme to cleave Zn-Sub. Then, two separated fragments of Zn-Sub respectively hybridize with the fluorescent probe and quencher probe, which results in a dramatic increase in the fluorescence intensity. The proposed aptasensor shows high sensitivity and selectivity for AFB1 detection with a detection limit of 0.22 nmol/L in a linear range of 0.4–16 nmol/L. Moreover, the fluorescent aptasensor exhibits acceptable applicability for detecting AFB1 in oil samples with satisfactory recoveries of 92.2–107.8%. Results are also in agreement with those of the ELISA method, indicating that the fluorescent sensing strategy has great potential applications in food safety control.
研究领域:
分析化学
·
生物技术
·
食品科学
·
分子生物学
✓ 已验证
DOI
PubMed
高影响因子
SENSORS AND ACTUATORS B-CHEMICAL
·
2018
·
IF 5.667
Li Zou,Xinghui Li,Tingting Li,Liansheng Ling
We have constructed a novel colorimetric sensing platform for quantitative detection of acetylcholinesterase (AChE) activity and its inhibitor (donepezil) in rat blood, which integrates the signal amplification of DNAzyme and hybridization chain reaction (HCR) with the assembly of gold nanoparticles (AuNPs). Herein, in the presence of AChE and its substrate acetylthiocholine (ATCh), the sensing system exhibits a dramatic decrease in absorbance at 522 nm, where AChE hydrolyzes ATCh into thiocholine, the resulting thiols capture Cu 2+ from DNAzyme, and then the substrate strand of DNAzyme acts as an initiator to trigger HCR process. The HCR products can induce the assembly of AuNPs via DNA hybridization, accompanying by a sharp color-change from red to blue. When donepezil is added, the enzymatic activity of AChE is suppressed, resulting in an increase in the absorbance at 522 nm. Under optimal conditions, the colorimetric sensing platform shows sensitive responses to AChE and donepezil in the range of 0.02–1.5 mU mL −1 and 0.5–100 nM, respectively. The detection limits of AChE and donepezil are as low as 5 μU mL −1 and 0.5 nM, respectively. Owing to high sensitivity of the proposed method, various nonspecific interactions can be eliminated with a sufficient dilution, indicating that our strategy has great potential for practical application in neurobiology and pharmacology fields.
研究领域:
生物化学
·
分子诊断
·
药物筛选
✓ 已验证
DOI
高影响因子
BIOELECTROCHEMISTRY
·
2020
·
IF 4.722
Hui Wang,Qingyao Luo,Yiguang Zhao,Xuemei Nan,Fan Zhang,Yaping Wang,Yue Wang,Dengke Hua,Shanshan Zheng,Linshu Jiang,Liang Yang,Benhai Xiong
Calcium is one of the most abundant and indispensable elements in biology, as it is a vital component of nerves, bones, and muscles and maintains the excitability of normal neuromuscular muscles. However, it may be harmful to the human body and even damage the organs if the calcium content exceeds the standard value by several times. To evaluate the level of calcium ions (Ca 2+ ), an electrochemical biosensor (FET/SWNTs/Cazyme) was developed using a nonspecific DNAzyme with high stability, which combined the unique advantage of field-effect transistors and single-walled carbon nanotubes, while being easy-to-use and having excellent sensitivity. The incubation time and voltage after optimization were 15 min and +0.02 V. The nonspecific DNAzyme-based biosensor was sensitive to Ca 2+ , but it was also interfered with by Pb 2+ , which affected the detection accuracy. To solve this shortcoming, an electrochemical device was proposed, in which FET/SWNTs/Cazyme combined with other specific biosensors for Pb 2+ , and then established some data processing models were established through support vector machine regression (SVMR) and artificial neural network fitting (ANNF). For the optimal SVMR, the electrochemical device can determine the Ca 2+ concentration in the range of 7.5–1000 μM with a detection limit of 5.48 μM. Finally, the prepared electrochemical device was employed to detect the Ca 2+ in different milk and water samples.
研究领域:
生物化学
·
分析化学
·
医学检测
✓ 已验证
产品关联
DOI
PubMed
高影响因子
ANALYTICA CHIMICA ACTA
·
2025
·
IF 6
Xin Meng,Yuanyuan Yue,Mengyang Huang,Zhangqun Duan,Kang Liu,Lina Wu
Background Lead ion (Pb 2+ ) is a toxic heavy metal that poses severe threats to food safety. Traditional methods like inductively coupled plasma mass spectrometry (ICP-MS) and atomic absorption spectrometry (AAS) rely on bulky instrumentation, which are limited by high costs, complex sample preparation requirements, and inability to meet the demands for rapid on-site testing. Nanozyme-based biosensors have emerged as promising alternatives, yet single-mode sensors often suffer from matrix interference in complex food samples. Therefore, developing a rapid and reliable on-site method is critical for Pb 2+ detection. Results To address this challenge, a CRISPR/Cas12a-driven dual-mode biosensor integrating Pt/CeO 2 nanozyme-mediated peroxidase activity with GR-5 DNAzyme recognition was developed. The biosensor's design capitalizes on GR-5 DNAzyme for Pb 2+ -specific recognition, triggering CRISPR/Cas12a-mediated cleavage of electrochemical/colorimetric signal probe SH-ssDNA-Pt/CeO 2 to generate dual signals. This innovative platform synergizes electrochemical precision and colorimetric simplicity for Pb 2+ detection in complex food matrices. The biosensor achieved an ultra-sensitive electrochemical response (linear range: 0.002–200 nM; limit of detection: 0.14 pM) alongside a robust colorimetric readout (linear range: 0.5–2000 nM; limit of detection: 0.47 nM), representing significant sensitivity improvements over conventional single-mode sensors. Crucially, the intrinsic cross-verification mechanism between orthogonal signal modalities minimized false positives while ensuring >90.5 % recovery in spiked corn, edible oil, beef and red wine samples with RSD <5 %. These results highlight the potential of the dual-mode sensor as a practical, field deployable sensing platform for the detection of Pb 2+ . Significance This work innovatively integrates the specificity of CRISPR with the catalytic properties of nanozymes into a self-validating electrochemical/colorimetric dual-mode system. The detection method not only establishes a robust platform for highly sensitive, reliable and visualized detection of Pb 2+ in complex food matrices, but also pioneers a new paradigm for multi-modal biosensor design.
研究领域:
分析化学
·
食品安全
·
生物传感器
·
环境毒理学
·
分子诊断
·
纳米生物技术
✓ 已验证
产品关联
DOI
PubMed
高影响因子
Analytical Methods
·
2022
·Vol. 14
·期 44
·4523-4530
·
IF 3.532
Jiao Wang,Liyuan Wen,Ruyuan Cao,Xiaorong Gao,Xia Li,Ensheng Xu,Qi Zhang,Shuling Xu,Caifeng Dai,Qingwang Xue
Single miRNAs are not specific and accurate enough to meet the strict diagnosis requirements in practice. Therefore, simultaneous monitoring of multiplexed miRNA in biological samples can not only improve the accuracy and specificity of bioassays but also avoid the squandering of valuable biological specimens. Herein, we designed a metal sulfide nanoparticle-based dual barcode-triggered DNAzyme cascade strategy for the sensitive and simultaneous multiplex miRNA detection in a single assay. Firstly, the capture probes (H1, H2) specifically recognize targets (miRNA-21, miRNA-141), exposing the stem of H1 and H2. Then, with the introduction of a detection probe (CuS-H3, ZnS-H4), the exposed H1 and H2 catalyze the hairpin assembly (CHA) reaction, realizing target miRNA recycling, and forming H1/H3-CuS and H2/H4-ZnS complexes. Subsequently, the formed H1/H3-CuS and H2/H4-ZnS complexes are encoded on magnetic beads through the biotin/streptavidin interaction. The CuS and ZnS nanoparticles captured by magnetic beads release thousands of Cu2+ and Zn2+via the cation exchange reaction. Finally, the released Cu2+ and Zn2+ specially activate the DNAzyme of the catalytic and molecular beacon (CAMB) system. The CAMB system affords an amplified fluorescence signal output by cycling and regenerating the metal ion-dependent DNAzyme to realize multiple enzymatic turnovers. Benefiting from target recycling, nanoparticle amplification, and catalytic and molecular beacon amplification, there is substantial amplification and the target miRNAs can be detected at 0.06 fM (miRNA-21) and 0.048 fM (miRNA-141) in a single assay. Furthermore, the high selectivity and accuracy of the assay were proved by practical analysis of different cancer cells, which exhibited good practicability in multiplex miRNA detection in clinical sera. The results indicate that the proposed strategy holds great potential for the sensitive detection of multiplex cancer biomarkers and offers the opportunity for future applications in clinical diagnosis.
研究领域:
生物化学
·
分子诊断
·
纳米医学
✓ 已验证
产品关联
DOI
PubMed
高影响因子
International Journal of Nanomedicine
·
2023
·
IF 8
Jiebing Yang,Jiayuan Zhang,Jiakai Xing,Zhiyuan Shi,Haobo Han,Quanshun Li
Background The phenylboronic acid-functionalized polyamidoamine (PP) was employed as a gene carrier for Dz13 delivery, inducing an obvious anticancer response.Materials and methods The Dz13 condensation ability of PP was evaluated through gel retardation assay. The cellular uptake mechanism of PP/Dz13 nanoparticles was studied using confocal laser scanning microscope and flow cytometer. The inhibition ability of cell proliferation, migration and invasion was investigated through MTT assay, flow cytometry, wound healing and Transwell migration assays, using hepatocarcinoma cell line HepG2 as a model. Finally, Western blotting analysis was used to detect the signaling pathway associated with the inhibition of cell apoptosis and migration induced by Dz13 delivery.Results The carrier PP could efficiently condense Dz13 into stable nanoparticles at mass ratios of >1.5. The hydrodynamic diameter and zeta potential of PP/Dz13 nanoparticles were measured to be 204.77 nm and +22.00 mV at a mass ratio of 10.0, respectively. The nanoparticles could realize an efficient cellular uptake in sialic acid-dependent endocytosis manner. Moreover, the nanoparticles exhibited an obvious antiproliferation effect through the induction of cell apoptosis and cell cycle arrest due to the cleavage of c-Jun mRNA. Besides, the suppression of cell migration and invasion could be achieved after the PP/Dz13 transfection, attributing to the decreased expression level of MMP-2 and MMP-9.Conclusion The PP provided a potential delivery system to achieve the tumor-targeting gene therapy.
研究领域:
分子生物学
·
基因治疗
·
肿瘤学
✓ 已验证
产品关联
DOI
PubMed
高影响因子
Nanoscale
·
2015
·Vol. 7
·期 19
·9055-9061
·
IF 7.394
Wenjiao Zhou,Wenbin Liang,Xin Li,Yaqin Chai,Ruo Yuan,Yun Xiang
The construction of DNA nanostructures with various sizes and shapes has significantly advanced during the past three decades, yet the application of these DNA nanostructures for solving real problems is still in the early stage. On the basis of microRNA-triggered, catalytic self-assembly formation of the functional “DNAzyme ferris wheel” nanostructures, we show here a new signal amplification platform for highly sensitive, label-free and non-enzyme colorimetric detection of a small number of human prostate cancer cells. The microRNA (miR-141), which is catalytically recycled and reused, triggers isothermal self-assembly of a pre-designed, G-quadruplex sequence containing hairpin DNAs into “DNAzyme ferris wheel”-like nanostructures (in association with hemin) with horseradish peroxidase mimicking activity. These DNAzyme nanostructures catalyze an intensified color transition of the probe solution for highly sensitive detection of miR-141 down to 0.5 pM with the naked eye, and the monitoring of as low as 283 human prostate cancer cells can also, theoretically, be achieved in a colorimetric approach. The work demonstrated here thus offers new opportunities for the construction of functional DNA nanostructures and for the application of these DNA nanostructures as an effective signal amplification means in the sensitive detection of nucleic acid biomarkers.
研究领域:
生物化学
·
分子诊断
·
纳米技术
·
癌症研究
✓ 已验证
DOI
PubMed
高影响因子
ANALYST
·
2019
·Vol. 144
·期 6
·1948-1954
·
IF 4.019
Xin Gong,Xiao Li,Taiping Qing,Peng Zhang,Bo Feng
Tetracycline (TC) is widely used to treat bacterial infections in humans and animals due to its low price and good antibacterial properties. The abuse of tetracycline has led to TC residues in daily food that could seriously affect human health. Thus, it is imperative to develop highly sensitive and selective methods for TC detection. In this work, we developed a colorimetric method for TC detection based on an enzyme-linked aptamer assay (ELAA) with multivalent HRP-mimicking DNAzyme. An aptamer was used as an alternative recognition element in the enzyme-linked immunosorbent assay (ELISA). Multivalent HRP-mimicking DNAzyme, assembled via hybridization chain reactions (HCR), was used for catalytic substrate color rendering in ELAA. The multivalent HRP-mimicking DNAzyme exhibited enhanced catalytic capacity and improved the detection sensitivity greatly. The limit of detection was 8.1 × 10−2 ng mL−1 with a linear range from 1.0 × 10−2 ng mL−1 to 1.0 × 104 ng mL−1 toward TC in buffer. To challenge the practical application capability of this strategy, the detection of TC in milk samples was also investigated and showed similar linear relationships. Due to the introduction of an aptamer, this ELAA strategy shows high selectivity towards TC and has potential for the detection of a wide spectrum of analytes.
研究领域:
分析化学
·
食品安全
·
生物技术
✓ 已验证
产品关联
DOI
PubMed
高影响因子
NEW JOURNAL OF CHEMISTRY
·
2022
·Vol. 46
·期 21
·10394-10401
·
IF 3.925
Jingjing Li,Long Jiang,Xu Wang,Zhixue Zhu,Qingxin Zhang,Su Liu,Yu Wang,Jiadong Huang
Kanamycin residue has attracted much attention due to its potential risk to the environment and human health. Herein, an electrochemical platform with superior performance was proposed based on enzyme-assisted target recycling with a strand displacement amplification (SDA) reaction and a DNAzyme-derived DNA walker. Our sensing system consists of an SDA amplification system and a DNA walker system. In the DNA walker system, a DNAzyme walker (DW)–locking probe (LP) and signal hairpin (HP) were assembled on the surface of a gold electrode (AuE) through a gold sulfur bond (Au–S). The specific binding of kanamycin with its aptamer induced an enzyme-assisted SDA process, leading to the generation of output probes (OP) which can open the locked DW-LP. And then, in the presence of Mn2+, the unlocked DW walked along the HP on the electrode surface cutting the loop of HP, so that the HP was consumed to form a G-quadruplex (G4) in the presence of K+. G4/hemin has peroxidase-like activity and can catalyze the reduction of H2O2 to produce obvious electrical signals. Upon the optimization of the analytical conditions, the proposed biosensor showed a wide linear range from 100 fM–5 nM with a limit of detection (LOD) of 87.1 fM. In addition, the aptasensor also had good stability and specificity, showing good prospects in practical applications.
研究领域:
生物化学
·
分析化学
·
环境科学
✓ 已验证
产品关联
DOI
高影响因子
SENSORS AND ACTUATORS B-CHEMICAL
·
2017
·
IF 5.401
Xu Hun,Yan Meng,Shiyu Wang,Zhenghua Mei,Xiliang Luo
A nonlinear hybridization chain reaction (non L-HCR) is presented here, in which series of toehold-mediated strands hybridization and branch migration reaction initiate in the presence of a trigger DNA. It leads to self-sustained assembly of double-stranded substrates into fractal nanostructures. The formed fractal nanostructures were about 10 μm length × 1.5 μm wide × 30 nm high. Furthermore, the 3′ part (TAGGGTTGGGCGGGATGGGT) of Gx DNA are exposed and ready to combined with hemin to form hemin/G-quadruplex DNAzyme, which could amplify the generation of chemiluminescence (CL) in the presence of luminol and H 2 O 2 . Under optimal conditions, the CL intensity linearly increased with the concentration of the trigger DNA from 0.5 × 10 −6 to 1.0 × 10 −11 M and the detection limit is 7.0 × 10 −12 M. In this assay, three-input concatenated logic gates operating as a biocomputing keypad-lock security system also have been successfully constructed. When the inputs are all introduced the “lock” opens. This simple non L-HCR design realizes the fractal nanostructures construction, CL detection and logic gates operating. It can be used in the nanotechnology and biological assay.
研究领域:
生物化学
·
纳米生物学
·
分析化学
✓ 已验证
产品关联
DOI
高影响因子
危险品化学品经营许可证（带存储）