ACS Sensors
·
2025
·
IF 9.1
Yapeng Wu,Bei Lv,Xintian Ni,Sheng Zhu,Dawei Li
Strand displacement amplification (SDA) is an isothermal DNA amplification technique. Herein, we developed a novel SDA system, designated All-U-Want SDA (AUW-SDA), which was used as a signal amplification strategy for the construction of nucleic acid detection biosensors. AUW-SDA is capable of target turnover and can be utilized for detection of nucleic acid sequences without available 3′-ends. Of particular significance is the ability of AUW-SDA to generate a substantial number of programmable sequences in accordance with the specifications of the sensor signal output methods, irrespective of the sequence of the target nucleic acid. We used the N gene of SARS-CoV-2 as a model target to develop a sensing platform with dual signal outputs. The colorimetric signals were generated by the G-quadruplex/hemin DNAzyme, in which the G-rich sequences were produced by AUW-SDA with a C-rich primer. On the other hand, by altering the sequence within the replaceable region of the primer, an activator sequence was obtained from AUW-SDA, which could trigger the activity of CRISPR/Cas12a, cleaving the probes modified with a fluorophore and quencher at each end and subsequently yielding the fluorescent signals. After the DNA sequences and reaction conditions were optimized, the limit of detection (LOD) values of the fluorescent and colorimetric assays were estimated to be 0.672 fM and 13.3 fM, respectively. The biosensors were utilized for biological sample detection. The reliability of the proposed method was validated against RT-qPCR results. In addition, a portable scanner-assisted high-throughput RGB analysis (PSHRA) method was developed. This method was applied to our biosensor for multilocus detection of SARS-CoV-2. The results obtained were satisfactory, indicating the potential of this approach for field testing or point-of-care (POC) diagnostics.
研究领域:
生物化学
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分子生物学
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诊断学
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DOI
PubMed
高影响因子
BIOSENSORS & BIOELECTRONICS
·
2014
·
IF 6.451
Xiaojun Chen,Lingna Ge,Buhua Guo,Ming Yan,Ning Hao,Lin Xu
An ultrasensitive electrochemical aptasensor was successfully fabricated for the detection of adenosine triphosphate (ATP). For the first time, one detection system combined several elements: magnetic aptamer sequences for target recognition and separation, a DNAzyme assisted cyclic signal amplification strategy, layer-by-layer (LBL) quantum dots (QDs) composites for promoting square wave anodic stripping voltammetric (SWASV) analysis and Bi, Nafion (Nf) and three-dimensional ordered macroporous polyaniline-ionic liquid (Bi/Nf/3DOM PANI-IL) film modified glassy carbon electrode (GCE) for monitoring enhanced SWASV signal. The modification of Nf/3DOM PANI-IL on GCE showed that the preconcentration efficiency was improved by the electrostatic absorption of Cd 2+ with negative Nf layer with the enhanced analytical sensitivity due to a large active surface area of 3DOM structure. The increased SWASV peak current values of the label (CdS) 4 @SiO 2 composites were found to be proportional to the logarithmic value of ATP concentrations in the range of 1 pM–10 nM and 10 nM–1 µM, with the detection limit as low as 0.5 pM. The proposed aptasensor has shown an excellent performance such as high sensitivity, good selectivity and analytical application in real samples. The results demonstrated that the multiple signal amplified strategy we developed was feasible for clinical ATP assay and would provide a promising model for the detection of other small molecules.
研究领域:
生物化学
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分析化学
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DOI
PubMed
高影响因子
ANALYTICAL AND BIOANALYTICAL CHEMISTRY
·
2018
·Vol. 410
·期 17
·4227-4234
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IF 3.307
Liu Hongwen,Chen Yating,Song Chunli,Tian Gang,Li Shiya,Yang Guiying,Lv Changyin
Radioactive radon decays into a stable daughter product, 210 Pb, which was used as the detection target to determine the radon radiation dose in a new technique. Pb 2+ triggers DNAzyme to cleave a molecular beacon (MB), resulting in the stem–loop structure opening and forming two single DNA strands (ssDNA). The ssDNA binds to unmodified gold nanoparticles and effectively prevents their aggregation in a salt solution. The detached enzyme strands continue to complement the remaining MB to amplify the response signal. The method proposed in this study exhibited a good linear relationship for Pb 2+ and radon concentrations in the range of 6.22 × 10 2 –1.02 × 10 5 Bq h/m 3 with a detection limit of 186.48 Bq h/m 3 using an ultraviolet–visible spectrometer. In practical applications, this sensitive method can avoid radioactive damage in field testing, and the detection limit meets the national standard in China. Importantly, this simple, highly sensitive strategy uses simple equipment and has a strong anti-interference ability.
Graphical abstract
研究领域:
辐射检测
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环境科学
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分析化学
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产品关联
DOI
PubMed
高影响因子
BIOSENSORS & BIOELECTRONICS
·
2015
·
IF 6.409
Ying Yang,Zheng Yuan,Xing-Pei Liu,Qiao Liu,Chang-Jie Mao,He-Lin Niu,Bao-Kang Jin,Sheng-Yi Zhang
The detection and speciation analysis of metal-ion is very important for environmental monitoring. A novel electrochemical biosensor for Nickel(II) detection based on a DNAzyme-CdSe nanocomposite was developed. We firstly hybridized with capture probe (DNA1) and sequentially with DNA (DNA2) on the gold electrode. Then CdSe QDs were incorporated the specific recognition of DNA2 by covalent assembling. Upon addition of nickel ion into the above system, the substrate strand of the immobilized DNAzyme was catalytically cleaved by target Ni 2+ , resulting in disassociation of the shorter DNA fragments containing CdSe QDs. The remaining CdSe QDs on the electrode surface detected by differential pulse anodic stripping voltammetry (DPASV). Under optimal conditions, the as-prepared sensor exhibited high sensitivity and fast response to Ni 2+ with the linear range from 20 nM to 0.2 mM and a low detection limit of 6.67 nM. The prepared biosensor also shows good stability and good reproducibility and high selectivity toward target Ni 2+ against other metal ions because of highly specific Ni 2+ -dependent DNAzyme. Thus, our strategy has a good potential in the environment surveys.
研究领域:
环境科学
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生物化学
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分析化学
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产品关联
DOI
PubMed
高影响因子
Chemical Science
·
2020
·Vol. 11
·期 33
·8966-8972
·
IF 9.346
Xia Li,Fang Yang,Wenjiao Zhou,Ruo Yuan,Yun Xiang
DNAzymes exhibit high potential as gene silencing agents for therapeutic applications. Such purposes, however, are significantly challenged by the targeted and successful delivery of unmodified DNAzymes into cells with minimal side effects. Here, we set out to formulate and demonstrate a new stimuli-responsive and constrained aptamer/DNAzyme (Apt/Dz) catenane nanostructure for highly specific gene silencing. The rational design of the Apt/Dz catenane nanostructure with the respective integration of the aptamer sequence and the completely closed catenane format enables both the targeted capability and significantly improved nuclease resistance, facilitating the stable and targeted delivery of unmodified Dz into cancer cells. Moreover, the Dz enzymatic activity in the constrained structure can only be conditionally regulated by the specific intracellular mRNA sequences to silence the target gene with highly reduced side effects. Results show that the Apt/Dz catenane nanostructure can effectively inhibit the expression of the target gene and the proliferation of cancer cells with high specificity.
研究领域:
分子生物学
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治疗学
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纳米技术
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癌症研究
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产品关联
DOI
PubMed
高影响因子
BIOSENSORS & BIOELECTRONICS
·
2021
·
IF 10.618
Wan Huang,Danyan Zhan,Yiming Xie,Xin Li,Guosong Lai
DNA walkers have been recognized as a type of powerful signal amplification tool for biosensors, but how to adopt a proper strategy to increase their amplification efficiency is still highly desirable. Herein we design a dual-catalytic hairpin assembly (CHA)-mediated strategy for the high-efficient formation of a tripedal Mg 2+ -dependent DNAzyme (MNAzyme)-DNA walker, and thus develop a novel proteinase-free dual-mode biosensing method for the kanamycin (Kana) antibiotic assay. The first CHA is initiated by a target-biorecognition reaction, which can produce the DNA walker and also induce the target recycling. The second CHA is initiated by a special base sequence designed as a one-half substrate of the MNAzyme. Upon the first CHA-triggered DNA walking at a magnetic bead (MB) track, this “pseudo-target” sequence can be released to induce another CHA-cycle for the formation of the same DNA walker. Meanwhile, the other one-half substrate strand exposed on the MB surface will trigger the quantitative hybridization chain reaction (HCR)-assembly of a G-quadruplex DNAzyme (G-DNAzyme)-enriched double-stranded DNA polymer. So the enzymatic reaction of G-DNAzymes enabled the convenient colorimetric and photoelectrochemical dual-mode signal transduction of the method. Due to the dual-CHA facilitation to the tripedal and three-dimensional DNA walking and synergetic signal amplification of HCR, this method exhibits very low detection limits of 9.4 and 0.55 fg mL −1 , respectively. In combination with its wide linear range, automated manipulation, and excellent selectivity , repeatability and reliability, the proposed method is expected to be used for the convenient semiquantitative screening and accurate determination of possible antibiotic residues in complicated matrices.
研究领域:
生物化学
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分析化学
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生物传感器
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纳米技术
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产品关联
DOI
PubMed
高影响因子
SENSORS AND ACTUATORS B-CHEMICAL
·
2018
·
IF 5.667
Liaojing Huang,Liu Yang,ChuanChao Zhu,Hanmei Deng,Guangpeng Liu,Yali Yuan
It is of great significance to explore reliable and sensitive methods for the trace detection of lead ions (Pb 2+ ), which may cause serious adverse effect to human health. Herein, we proposed a methylene blue (MB) sensitized signal-on photoelectrochemical (PEC) biosensor with 3,4,9,10-perylene tetracarboxylic acid (PTCA) film as photoelectric material for highly sensitive Pb 2+ detection. First, PTCA was modified on the electrode surface, manifesting wonderful film-forming property and acquiring a satisfactory initial PEC signal. Afterward, Pb 2+ -dependent DNAzyme modified on electrode was cleaved by target Pb 2+ , followed by hybridizing MB assembled 3D DNA networks with the exposed Pb 2+ catalytic strand. Significantly, in the presence of sensitizer MB, the PEC signal of PTCA was extraordinarily increased and the designed biosensor performed a high sensitivity with a detection limit down to 0.03 pM. Therefore, with enzyme-free assembly of MB embedded 3D DNA networks, this dye-sensitized strategy exhibited effective and uncomplicated amplification technology for Pb 2+ detection.
研究领域:
分析化学
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环境科学
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生物传感技术
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产品关联
DOI
高影响因子
COLLOIDS AND SURFACES B-BIOINTERFACES
·
2018
·
IF 3.997
Hai-Bo Wang,Li-Hong Ma,Bi-Yun Fang,Yuan-Di Zhao,Xue-Bin Hu
This study has reported that a GR-5 DNAzyme based lead ion strip biosensor could exhibit an enhanced specificity with the assistance of graphene oxide (GO). This enhancement results from the specific π-stacking interaction between the ribose rings of the nucleobases and the carbon hexagons in GO which can reduce the false positive interference by removing unhybridized ssDNA during the annealing of GR-5 DNAzyme. Meanwhile, conjugate pad was sprayed with two kinds of AuNP-DNA probes, and nitrocellulose membrane test zone and control zone were immobilized with two kinds of biotin-DNA probes, respectively. The limit of detection of this strip biosensor was estimated to be about 0.05 nM (S/N = 3) and 1 nM (with naked eyes) with a linear range from 0.01 to 100 μM. Furthermore, the strip biosensor exhibited excellent selectivity toward Pb 2+ in the presence of other divalent metal ions. For real soil samples, the obtained recoveries were in the range from 91.5% to 113.1%.
研究领域:
生物传感技术
·
环境科学
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产品关联
DOI
PubMed
高影响因子
POLYMER
·
2022
·
IF 4.432
Cheng Deng,Hai Yang,Simin Liu,Zhiyong Zhao
Nanomaterials self-assembled by DNA amphiphiles have great potential application in catalysis, bioimaging and biomedicine. Here, we constructed two kinds of DNA-dendrimer amphiphiles including the G-quadruplex-dendrimer hybrid (G20-Den) and single-stranded DNA-dendrimer hybrid (D20-Den) as a control. Both of them self-assembled into nanofibers with a length of several micrometers in MES buffer, and the diameter of G20-Den nanofibers was smaller than that of D20-Den nanofibers due to the compact stacking of G-quadruplex formed in G20-Den hybrids. At the same time, the catalytic properties of G20-Den nanofiber/hemin, as horseradish peroxidase-mimicking DNAzymes, were investigated. Our results showed that G20-Den nanofiber/hemin, as well as the single-stranded G20/hemin system, exhibited higher catalytic peroxidation activity than D20-Den nanofiber/hemin. Furthermore, through simple ultrafiltration, the recycle of these nanofibers can be simply achieved. This research has provided a new solution for the construction of functional DNA nanomaterials.
研究领域:
生物化学
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纳米医学
·
材料科学
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产品关联
DOI
高影响因子
Analytical Methods
·
2023
·Vol. 15
·期 37
·4972-4979
·
IF 3.1
Haiquan Liu,Yue Zhang,Weiqing Xiong,Xiaoyan Wang
Lead ion (Pb2+) detection is critically important in environmental protection and health management. In this work, we developed a simple signal-enhanced colorimetric sensor for the detection of Pb2+ based on the peroxidase-mimetic property of gold nanoparticles (AuNPs). When a certain concentration of Pb2+ was added to a solution of DNAzyme-modified AuNPs, aggregation was triggered, and the result was an enhancement of the peroxidase-mimetic activity of AuNPs. Then, the chromogenic reaction of 3,3′,5,5′-tetramethylbenzidine (TMB) by the catalyst of AuNPs was used for the sensitive UV-Vis and colorimetric detection of Pb2+. When a higher concentration of Pb2+ was added, the greater amount of aggregation of AuNPs resulted in the enhancement of the UV-Vis adsorption of the solution at 652 nm, with a deepening of the blue color of the solution. After optimization of the experimental conditions, a linear relationship between the absorbance of oxidized TMB at 652 nm and the logarithm of Pb2+ concentration was obtained, which had been divided into two parts (25 pM to 2.5 μM, and 2.5 μM to 250 μM). The detection limit was as low as 10 pM. The satisfactory specificity and rapid response of the sensor showed that it has promising application for the detection of Pb2+ in real samples.
研究领域:
环境科学
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纳米生物学
·
分析化学
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产品关联
DOI
PubMed
高影响因子
CHEMICAL ENGINEERING JOURNAL
·
2024
·
IF 13.3
Zhan Huang,Xing Huang,Yanfei Zhang,Meng Chen,Shaoqiang Wen,Zhou Zhang,Zong Dai,Xiaoyong Zou,Si-Yang Liu
DNAzyme-based gene silencing is a promising cancer therapy technology, while its efficiency and safety issues still need to be addressed. In this study, we developed a chirality/microRNA dual-gating theranostic nanomachine for messenger RNA (mRNA) gene silencing therapy with enhanced targeting towards human cervical cancer (HeLa) cells. The chiral nanoparticle, an emerging material with unique chirality-induced properties, e.g. chirality-dependent cellular uptake, has been widely applied in biomedical fields. The L-type gold nanoparticles (L-AuNPs) are used as the nanocarrier with the first “chirality-gating”. L-AuNPs showed 2.4-fold uptake efficiency for HeLa cells comparing to normal cell, as well as 2.2-fold uptake efficiency in cancer cell comparing to D-type gold nanoparticles (D-AuNPs), which exhibited no cancer cell targeting effect. Moreover, an entropy-driven-DNAzyme (EDz) circuit that can release numerous DNAzyme and fluorescence probes with the trigger of single microRNA (miRNA) conducted the theranostic role of both miRNA imaging and gene silencing, constituting the second “miRNA gating”. Ascribing to the prominent targeting nanocarrier L-AuNPs, the miRNA imaging sensitivity and gene silencing therapy efficiency were improved by 2.6- and 5.0-fold, respectively, comparing to those by D-AuNPs. In general, this dual-gating nanomachine, designed with a powerful yet simple strategy, has showed great prospects in optimizing delivery specificity and avoiding off-target toxicities.
研究领域:
癌症治疗
·
纳米技术
·
分子生物学
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生物医学工程
·
基因治疗
✓ 已验证
产品关联
DOI
高影响因子
MICROCHEMICAL JOURNAL
·
2021
·
IF 4.821
Xiaojie Song,Man-Man Lv,Qi-Yan Lv,Hui-Fang Cui,Jing Fu,Yu-Yang Huo
A novel and sensitive Helicobacter pylori assay strategy based on linear isothermal amplification reaction (LIAR) and DNAzymes catalyzed DNA sensor was developed. A 17 nt target DNA (tDNA) specific in the H. pylori genome was selected and used as primer in the LIAR. A template DNA was designed to produce an amplicon with Mg 2+ -dependent and RNA-cleaving DNAzyme (Md-Dz) activity. The amplicon (i.e. Md-Dz) was then detected with the DNA sensor. The DNA sensor was fabricated simply through immobilization of a hairpin DNA that contains both Md-Dz substrate and G-quadruplex DNAzyme (Gq-Dz), on gold electrodes. The Gq-Dz possesses peroxidase activity, catalyzing the electro-redox of the electrochemical signal probe hydroquinon. The LIAR, Md-Dz catalytic reaction, and DNA sensor fabrication and detection processes were characterized and optimized. The LIAR amplification efficiency is as high as ~5 × 10 5 fold, and the assay strategy is very sensitive, selective and repetitive. The differential pulse voltammetry (DPV) detection signals linearly correlate to the logarithmic tDNA concentration in the range of 0.1–10,000 fM, with a limit of detection (LOD) of 34 aM. The detection linear range for H. pylori genomic DNA is 2.1–67.2 pg, with an LOD of 1.3 pg . Muted DNA sequences and other pathogenic bacteria did not result in interferences. In addition, the DNA sensor is very stable in storage. More importantly, the DNA sensor and the assay strategy could be universally applicable to detect any biological sample and short DNA sequence of interests.
研究领域:
生物化学
·
分析化学
·
分子生物学
·
生物技术
✓ 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）