BIOELECTROCHEMISTRY
·
2022
·
IF 5.76
Hao Fan,Ying Wu,Tongfu Huang,Nian Hong,Hanfeng Cui,Guobing Wei,Fusheng Liao,Jing Zhang
As an artificial nanomachine, a DNA walker demonstrates the potential for biosensing. In this study, a highly integrated, biostable, and autonomous electrochemical DNA walker sensor was rationally designed by a simple assembly of a Mn 2+ -dependent DNAzyme-powered DNA walker with nanoscale Mn 2+ @MOFs containing free carboxylic acid groups UiO-66(Zr)-(COOH) 2 . In this study, the release of Mn 2+ from Mn 2+ @MOFs was exploited to drive the autonomous and progressive operation of the DNA walker, and the DNAzyme-driven DNA walker was constructed by the co-modification of walking strands and track strands onto the gold electrode (GE) surface. The walking strand was a single-stranded DNA containing a DNAzyme sequence, which was pre-silenced by the locking strand. The track strand was a specially designed DNA sequence that the target can hybridize with the locking strand; hence, the walking strand is unlocked, and the liberated DNAzyme catalyzes the cleavage of track strands to drive the DNA walker operation, shifting tetraferrocene away from the electrode and producing a significant signal change. A detection limit of 38 fM was obtained with our new system, exhibiting a wide linear range from 1.5625 × 10 −9 M to 1 × 10 −13 M. The proposed approach provided a novel means for constructing an highly integrated, automated, and DNAzyme-driven DNA walker for bioanalysis.
研究领域:
生物传感
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纳米技术
·
分子生物学
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产品关联
DOI
PubMed
高影响因子
NEW JOURNAL OF CHEMISTRY
·
2023
·Vol. 47
·期 42
·19413-19420
·
IF 3.3
Zongqiang Li,Jingjing Li,Xu Wang,Qianru Li,Jingru Zhu,Yuying Yao,Zhiqiang Guo,Yu Wang,Su Liu,Jiadong Huang
Escherichia coli (E. coli) is a common Gram-negative bacterium that is widely distributed in nature and is susceptible to natural and random genetic factors. It is a pathogenic factor for many diseases. Sensitive and specific detection of E. coli is an important means to ensure food safety and prevent foodborne disease outbreaks. Ultrasensitive and specific detection of E. coli is still an urgent problem that needs to be solved. An intuitive and convenient colorimetric assay method with high sensitivity based on a DNAzyme-powered AuNR-DNA walker is proposed here for detection of E. coli. The strategy, dependent on the targeted activation of Mg2+-DNAzyme, starts an efficient DNA walking machine, induces the formation of countless peroxidase-mimicking enzymes, and produces a color change via a chromogenic reaction for visual detection of E. coli. This strategy features several aspects. First, this is the first time that an AuNR-based DNA walker is integrated with a colorimetric assay of E. coli. By combining the DNAzyme-powered AuNR-DNA walker and visual detection via the peroxidase-mimicking enzyme, the detection sensitivity is significantly improved compared to those of previously reported colorimetric-based methods, which provides a paradigm for the development of highly sensitive colorimetric assay methods. Second, an activity-controlled nano-DNAzyme is constructed via target-triggered formation of G-quadruplex DNAzyme to achieve low background and high signal-to-noise detection. In addition, this strategy has the advantages of simple operation, short analysis time, low cost, and easy on-site detection. Therefore, the proposed strategy has the potential for application in the detection of E. coli and related food safety analysis and clinical diagnosis.
研究领域:
生物化学
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分子生物学
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纳米技术
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产品关联
DOI
高影响因子
SPECTROCHIMICA ACTA PART A-MOLECULAR AND BIOMOLECULAR SPECTROSCOPY
·
2024
·
IF 4.3
Bowen Xiong,Xiang Li,Yuting Zhang,Changling Yan,Gongke Wang
The overexpression of vascular endothelial growth factor 165 (VEGF 165 ) in cancer cells plays a pivotal role in promoting tumor metastasis by facilitating their excessively rapid proliferation and division. Hence, the development of analytical methods possessing high sensitivity and resistance to interference is imperative for the detection of VEGF 165 . Various types of aptasensors have been devised for VEGF 165 detection; however, the performance of these biosensors can be influenced by non-target signals caused by conformational changes in unbound aptamers. The paper shows the creation of a precise and sensitive fluorescence biosensor designed to detect VEGF 165 by using a VEGF 165 -specific split aptamer. Additionally, this biosensor employs nicking enzyme-assisted DNA walker coupling with CRISPR-Cas12a to achieve dual-signal amplification. The VEGF 165 calibration curve shows a detection limit of 268 fM and has a broad linear range from 5 to 4000 nM. The fluorometric biosensor was utilized to detect VEGF 165 in human serum and cellular homogenate samples, yielding good outcomes. The innovative design serves as proof of concept and demonstrates significant potential in detecting various targets.
研究领域:
生物技术
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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
TALANTA
·
2019
·
IF 4.916
Longsheng Jin,Jingtang Qiao,Jinghan Chen,Ning Xu,Meisheng Wu
A novel area controllable biosensing interface is designed on glassy carbon bead (GCB) and used for measurement of tetracycline (TET) in closed bipolar electrode-electrochemiluminescence (ECL) device. One face of GCB is modified with Au particles and the covered area is varied from 0 to 45.3% by tuning the external voltage during bipolar electrodeposition process in a home-made open bipolar electrochemical cell . It enables the conjugation of various amounts of biomolecules on Au/GCB. Then DNA walker and methylene blue (MB) labeled DNA (MB-DNA) are conjugated on Au surface for the following combination of aptamer . In the presence of target, aptamer partially hybridized with DNA walker will be released from Au surface, leading to the formation of dsDNA between DNA walker and MB-DNA. As a result, MB-DNA with recognition site for nicking endonuclease (Nb. Bbv CI) in dsDNA is cleft into two segments by Nb. Bbv CI. Meanwhile, the liberated DNA walker is triggered to continue the degradation of many MB-DNA. Due to the excellent electrochemical performance of MB, it is reduced at the cathode of BPE to amplify the ECL signal at the anode of BPE in a closed BPE-ECL platform. When the covered area of GCB by Au particles is enlarged from 10.1 to 45.3%, the change of ECL intensity could be increased 2.7-fold. The linear range for the detection of tetracycline is from 1 × 10 −12 to 1 × 10 −5 M with the detection limit of 6.0 × 10 −13 M. Hence, controllable sensing area on a single glassy carbon bead, the amplification effect of DNA walker and MB make this approach possess many advantages over traditional biosensor.
研究领域:
分析化学
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生物传感技术
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产品关联
DOI
PubMed
高影响因子
TALANTA
·
2025
·
IF 6.1
Xiang Li,Yuting Zhang,Mengyuan He,Jiarui Sun,Bowen Xiong,Gongke Wang
In this work, we present an ultrasensitive, specific, and high-signal-to-background ratio fluorescence split-aptasensor for D-vasopressin (D-VP) detection. This sensor is based on target-induced self-propelled 3D DNA walkers in conjunction with CRISPR-Cas12a technology. Two split probes (SDA 1 and SDA 2) were designed to undergo structural recombination and function as a walking chain (SDA) under the induction of D-VP. Simultaneously, an intact Mg 2+ -dependent DNAzyme domain was formed at the tail of SDA and subsequently activated. The activated Mg 2+ -dependent DNAzyme continuously propelled the 3D DNA walker, enabling the generation of signal strand DNA (activator DNA). The activator DNA can subsequently trigger the activation of the Cas12a protein, enabling it to cleave the FAM-ssDNA-BHQ1 substrate. This process leads to signal amplification and the specific detection of D-VP. Under optimal conditions, the designed split-aptasensor exhibits excellent linearity across a concentration range of 5 ng/mL to 1215 ng/mL, with a detection limit (LOD) as low as 0.22 ng/mL. This split-aptasensor were employed to identify D-VP in human serum and urine samples, yielding highly satisfactory results. This unique design acts as a proof of concept and illustrates considerable promise for the detection of a wide range of analytes.
研究领域:
生物化学
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分子诊断
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纳米生物技术
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DOI
PubMed
高影响因子
TALANTA
·
2023
·
IF 6.1
Ying Liao,Yingqin Zhang,Aiwen Su,Yanli Zhang,Hongbin Wang,Wenrong Yang,Pengfei Pang
Protein kinase A (PKA) can regulate many cellular biological processes by phosphorylation substrate peptide or protein. Sensitive detection of PKA activity is critical for the PKA-related drug discovery and disease diagnosis. A new electrochemical biosensing method was developed for detection of PKA activity based on Zr 4+ -mediated DNAzyme-driven DNA walker signal amplification strategy. In this strategy, the special designed substrate peptide and a thiolated methylene blue-labeled hairpin DNA (MB-hpDNA) containing a single ribonucleic acid group (rA) could be anchored on the surface of gold electrode by Au–S bond. In the presence of adenosine triphosphate (ATP) and PKA, substrate peptide was phosphorylated and linked with walker DNA (WD) via the robust phosphate-Zr 4+ -phosphate chemistry. The linked WD hybridized with the loop region of MB-hpDNA to form a Mn 2+ -dependent deoxynuclease (DNAzyme), which cleaved the MB-hpDNA into MB-labeled fragment releasing away from electrode surface, resulting in a dramatic decrease of electrochemical signal and providing an electrochemical sensing platform for PKA activity detection. The response signal of the developed biosensor is proportional to the logarithm of PKA concentration in the range of 0.05 U mL −1 to 100 U mL −1 , with a detection limit of 0.017 U mL −1 at a signal to noise ratio of 3. Furthermore, the proposed method can also be applied for the evaluation of PKA inhibition and PKA activity assay in cell samples. Therefore, the proposed biosensor shows great promise as a universal tool for diagnostics and drug discovery of PKA-related diseases.
研究领域:
生物化学
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分析化学
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医学诊断
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产品关联
DOI
PubMed
高影响因子
BIOSENSORS & BIOELECTRONICS
·
2023
·
IF 12.6
Siqi Zhao,Yaxue Jia,Anping Wang,Jinlan Yang,Li Yang
We present a novel dual-mode DNA-walker based optical fiber biosensor (DMDW-Opt biosensor) for sensitive assay of micro-RNAs in bio-samples. In the sensor system , we develop a new strategy for the cascade amplification, DNA-walker/rolling cycle amplification (RCA), by the use of the residue track of the walker. The strategy can significantly improve the response of the sensor and avoid any tedious operation procedure. Dual-mode readouts, i.e., fluorescence and chemiluminescence , are measured independently without interfering with each other to achieve reliable and accurate analysis. Optical fibers with the surface modified by gold nanoparticles are utilized as the support for fabrication of the sensor, which would be benefit for developing miniaturized and portable sensing devices. The performance of the proposed method is evaluated by using micro-RNAs (MiR-155 and MiR-21) as the analytical target. The method is successfully applied for accurate determination of micoRNAs in human serum and MCF-7 cells. Our method can perform sensitive assays of MiR-155 with limit-of-detection as low as 97.72 fM and 11.22 fM, MiR-21 with limit-of-detection as low as 107.15 fM and 8.32 fM for the fluorescence- and the chemiluminescence-readout respectively, and the biosensor exhibits excellent specificity, reproducibility and storage stability, indicating its valuable potential applications for sensing trace-amount targets in complicated real samples.
研究领域:
生物传感
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纳米技术
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分子生物学
·
分析化学
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产品关联
DOI
PubMed
高影响因子
SPECTROCHIMICA ACTA PART A-MOLECULAR AND BIOMOLECULAR SPECTROSCOPY
·
2022
·
IF 4.831
Yan Zhou,Zefeng Wang,Shengnan Zhang,Le Deng
Integrating the advantages of catalytic hairpin components and multi-foot DNA walker, we designed a 16S rRNA detection probe template for pathogen bacteria, which utilizes DNA ligation quencher and dual catalytic [email protected] walker to induce signal recovery. The dual catalytic [email protected] walker uses the walking position of the target on the AuNP as a foothold to promote the reaction, so that the biosensing of the low-abundance target sequence can induce signal recovery. During the entire experiment, no enzyme is required, which can avoid the limitation of enzyme degradation under unfavorable conditions and the inability to detect the target. Most importantly, the detection template has the advantages of high sensitivity, and its detection limit is significantly better than that of single hairpin DNA walker probe. As the detection system can sensitively and rapidly detect its targeted bacteria and not rely on any enzyme and sophisticated instrumentation, it has great potential for sensitive and specific pathogenic bacteria detection.
研究领域:
分子生物学
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生物传感
·
病原体检测
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DOI
PubMed
高影响因子
Journal of Analysis and Testing
·
2022
·Vol. 6
·期 1
·12-19
·
IF 0
Wang Xin-Yan,Li Yuanfang,Lv Shuzhen,Bi Sai
Herein, a dual-signal electrochemical biosensor has been developed by self-assembly of pH-activatable i-motif probes on magnetic microparticles (MMPs) coupled with DNA walker for signal amplification. In this study, the cytosine (C)-rich single-stranded DNAs are hybridized with DNA initiators to obtain the long-nicked duplexes with repeated units, which are further captured on MMPs to form the magnetic i-motif containers. The resulting duplexes contain abundant G–C base pairs, thus providing extensive binding sites for doxorubicin (DOX). At acidic pH, the C-rich sequences are folded into i-motif structure, resulting in the release of DOX and walker initiators. In this case, the liberated DOX is adsorbed on graphene quantum dots-modified glassy carbon electrode via π–π interaction, while the walker initiators as a moving part can catalyze the hybridization between MB-modified fueler DNA and tracker DNA on electrode, contributing to the generation of dual electrochemical signals induced by MB and DOX. Importantly, the magnetic separation can effectively reduce the background, achieving sensitive biosensing of pH ranging from 4.0 to 7.4 with excellent stability. Moreover, the proposed dual-signal electrochemical biosensor has been successfully applied for accurate monitoring of pH in human serum, which holds great potential in pH-dependent bioassays, especially in ultra-micro analysis for clinical applications.
研究领域:
生物化学
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分析化学
·
医学诊断
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DOI
TALANTA
·
2022
·
IF 6.556
Tongfu Huang,Mei Wang,Nian Hong,Hanfeng Cui,Qiqi Fan,Guobing Wei,Longshua Qin,Jing Zhang,Hao Fan
Herein, an autonomous driven DNA walker-based aptasensor is proposed for the detection of Ochratoxin A (OTA). A nanoscale metal-organic framework (MOF) is synthesized and used to load Mn 2+ , a cofactor for DNAzyme. The DNA walker and Mn 2+ @MOF are assembled on the Au surface, integrating the DNA walker and metal auxiliary ions. The proposed sensor avoids the addition of metal auxiliary ions of DNAzyme from the external environment, which achieves the fully autonomous driving of DNA walker and facilitates the on-site detection of OTA. In addition, the substrate strands are modified with a newly synthesized tetraferrocene signal marker, further achieving signal amplification. The sensor demonstrates a sensitivity of 0.289 pg/mL and is successfully applied to real food sample analysis.
研究领域:
生物化学
·
分析化学
·
食品科学
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产品关联
DOI
PubMed
高影响因子
SENSORS AND ACTUATORS B-CHEMICAL
·
2023
·
IF 8.4
Jingyi Cai,Junping Ma,Yuan Dang,Shuangli Chen,Jieli Tang,Sha Yu,Yuanzhen Zhou
Antibiotics residue pollution in food and environmental matrices has posed a serious threat to global public health. It is an urgent demand in developing highly sensitive methods for the detection of low-abundance antibiotics in real samples. Herein, we established a magnetic DNA walker-engineered electrochemical sensor by taking advantages of the distinct recognition capability of aptamer and the excellent signal amplification performance of magnetic DNA walker for the accurate detection of kanamycin (Kana). The aptamer can specifically recognize Kana, while the magnetic DNA walker can not only amplify the target signal through the cascade displacement reaction, but also effectively reduce background interference via the magnetic separation, thus ensuring the sensitivity and accuracy of Kana detection in complex samples. The proposed electrochemical sensing method possesses a good linear relationship in the range of 10 fM −1 μM, and the limit of detection is as low as 0.02 fM. Moreover, the excellent selectivity allows this sensor has the ability to specifically identify the Kana from other antibiotics and even in the milk and tap water.
研究领域:
生物传感
·
环境科学
·
分析化学
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产品关联
DOI
高影响因子
RSC Advances
·
2022
·Vol. 12
·期 39
·25744-25752
·
IF 4.036
Wenzhuo Wang,Wei Yuan,Debao Wang,Xutao Mai,Daoying Wang,Yongzhi Zhu,Fang Liu,Zhilan Sun
Clostridium perfringens is an important foodborne pathogen, which has caused serious public health problems worldwide. So, there is an urgent need for rapid and ultrasensitive detection of C. perfringens. In this paper, a dual-mode sensing platform using the synergy between fluorescent and electrochemical signals for Clostridium perfringens detection was proposed. An electrochemical aptasensor was constructed by a dual-amplification technology based on a DNA walker and hybridization chain reaction (HCR). When the C. perfringens genomic DNA was present, it specifically bonded with FAM-labeled aptamer which triggered the DNA walker on hairpin DNA (hDNA) tracks to start the synthesis of double-stranded DNA. HCR occurred subsequently and produced long-chain DNA to absorb more methylene blue (MB). In this cycle, the fluorescent signals of released FAM-labeled aptamer could also be detected. The synergistic effects of MB and FAM significantly improved the sensitivity and accuracy of the dual-mode sensor. As a result, the biosensor displayed an excellent analytical performance for C. perfringens at a concentration of 1 to 108 CFU g−1. A minimum concentration of 1 CFU g−1 and good accuracy were detected in real samples. The proposed ultrasensitive detection method for detecting C. perfringens in food showed great potential in controlling foodborne diseases.
研究领域:
生物传感技术
·
食品安全检测
·
分子生物学
✓ 已验证
DOI
PubMed
高影响因子
危险品化学品经营许可证（带存储）