ANALYST
·
2025
·
IF 3.3
Daqi Chen,Zhezhi Fang,Yachunyue Zhou,Ziyao Zeng,Jiancong Liang,Baian Zhu,Chaozhan Chen
MicroRNAs (miRNAs), as pivotal regulators of gene expression, are critically associated with the progression of cancer, neurodegenerative, and cardiovascular diseases. However, their detection remains challenging due to low sensitivity, background noise, and matrix interferences in complex biological samples, hindering clinical applications such as early tumor screening and liquid biopsy. Herein, we present a cascade-amplified fluorescence biosensor driven by the synergistic integration of exonuclease III (Exo III) and self-locking DNAzyme (SLD). This strategy exploits Exo III-mediated cyclic digestion of target-induced DNA/RNA heteroduplexes to release multiple signal conversion chains, while the SLD’s rigid conformational gate suppresses nonspecific catalysis until unlocked by target-specific molecular keys. The activated DNAzyme then drives secondary amplification through iterative cleavage of fluorophore-labeled reporters. This dual-amplification mechanism achieves label-free, isothermal detection of miRNA-155 with a linear range of 50 fM–10 pM and a limit of detection (LOD) of 4.96 fM. Notably, the sensor demonstrates robust specificity in serum matrices, effectively overcoming homologous miRNA cross-reactivity and complex biological interferences. By merging target recycling with conformation-gated catalysis, this platform offers a cost-effective, high-precision solution for miRNA profiling, holding significant promise for point-of-care diagnostics and liquid biopsy-based early cancer detection.
研究领域:
分子生物学
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肿瘤学
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转化医学
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生物传感
·
临床检验
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DOI
PubMed
高影响因子
JOURNAL OF HAZARDOUS MATERIALS
·
2022
·
IF 14.224
Biaoliang Wu,Xiaoqing Wen,Jiuying Cui,Xiaojie Qin,Zheng Li,Yuanxun Gong,Qianli Tang,Xianjiu Liao,Zhenqiang Fan,Kai Zhang
Rapid and accurate discrimination of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) is an available approach to implement a rapid diagnosis of the coronavirus disease 2019 (COVID-19). Here we fully exploited the cleavage properties of exonuclease III (Exo III) and hairpin DNA-assisted target cycling technology to generate bulk single-stranded DNA (ssDNA) that was employed to facilitate the constitution of a three-way junction structure on polymetallic particle (Ag-Au NPs) and Ti 3 C 2 (Ti 3 C 2 @Ag-Au) complexes. Ag-Au NPs presented favorable stability without adding extra stabilizers, demonstrating the potential value of Ag-Au NPs as an alternative to Au NPs in the field of bioanalysis. Uppon the three-way junction structure, the dumbbell hybridization chain amplification (DHCA) was occurred which generated DNA nanostructure with tight conformation. Target cycling and DHCA reactions improved the electrochemiluminescence (ECL) signal, which dramatically advanced the assay sensitivity of SARS-CoV-2 (0.59 fM). Moreover, our strategy remained to demonstrate favorable specificity and repeatability in environmental conditions and real human serum samples.
研究领域:
生物分析
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纳米技术
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病毒学
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诊断技术
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DOI
高影响因子
INTERNATIONAL JOURNAL OF BIOLOGICAL MACROMOLECULES
·
2022
·
IF 8.025
Yufei Liu,Suyu Meng,Jingjing Qin,Ruiying Zhang,Ningning He,Yaoyao Jiang,Hong Chen,Na Li,Ying Zhao
As a toxic substance on human health produced in food thermal treatment, simple analytical approaches are highly desired for the detection of acrylamide (ACR) in foods. With the aid of exonuclease III (Exo III), a simple fluorescence sensor was proposed based on carboxyfluorescein-labeled double-stranded DNA (FAM-dsDNA) and a cationic conjugated polymer (PFP). Fluorescence resonance energy transfer (FRET) efficiency between FAM and PFP was changed with and without ACR. When ACR was present, ACR and single-stranded DNA (P1, ssDNA) formed an adduct, allowing free FAM-labeled complementarity strand DNA (P2, FAM-csDNA) to appear in the solution and avoiding the digestion of P2 by Exo III. After the addition of PFP, the interaction of PFP and FAM induced strong FRET. Under optimized conditions, ACR was detected with a limit of detection (LOD) of 0.16 μM. According to this biosensor, a LOD of 1.3 μM in water extract samples was observed with a good recovery rate (95–110 %).
研究领域:
食品科学
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分析化学
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生物技术
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DOI
PubMed
高影响因子
ANALYTICA CHIMICA ACTA
·
2020
·
IF 5.977
Dandan Huang,Jiaming Chen,Li Ding,Longhua Guo,Palanisamy Kannan,Fang Luo,Bin Qiu,Zhenyu Lin
In this work, core-satellite assemblies and exonuclease assisted double amplification strategy is developed to produce surface-enhanced Raman scattering (SERS) biosensor towards ultrasensitive detection of biotoxin. In the presence of target molecules, the exonuclease III (Exo III) assisted efficient recycling amplification provides an excellent pathway for the fabrication of core-satellite SERS sensor. Briefly, the proposed strategy includes the following double amplifications: (i) Exo III induced target-related signal amplification; (ii) core-satellite assemblies assisted formation of SERS “hot-spots” induced signal amplification. To show the applicability of the suggested strategy, the detection of ochratoxin A (OTA), one of the most toxic and widely distributed biotoxin, is demonstrated as an example. The results show that the limit of detection (LOD) of OTA is 0.83 fg mL −1 (S/N = 3). On the basis of the DNA aptamer induced specific target recognition, hence our sensing strategy is easy to be expended to the ultrasensitive detection of other targets, e.g., DNAs, RNAs, and other molecules that have corresponding DNA aptamers.
研究领域:
分析化学
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毒理学
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纳米技术
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生物技术
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DOI
PubMed
高影响因子
ANALYTICAL CHEMISTRY
·
2017
·
IF 6.32
Bingzhi Li,Lei Xu,Yue Chen,Wanying Zhu,Xin Shen,Chunhong Zhu,Jieping Luo,Xiaoxu Li,Junli Hong,Xuemin Zhou
Transcription factors (TFs) regulate gene expression by binding to regulatory regions, and their dysregulation is involved in numerous diseases. Thus, they are therapeutic targets and potential diagnostic markers. However, widely used methods for TFs detection are either cumbersome or costly. Herein, we first applied DNA-Ag nanoclusters molecular beacons (AgMBs) in TFs analysis and designed an assay based on the switchable fluorescence of AgMBs. In the absence of TFs, a single-stranded DNA functioned as a reporter is released from a double-stranded DNA probe (referred as dsTFs probe) under exonuclease III (Exo III) digestion. Then, the reporter triggers downstream Exo III-assisted signal amplification by continuously consuming the guanine-rich enhancer sequences in AgMBs, resulting in significant fluorescent decrease eventually. Conversely, the presence of TFs protects the dsTFs probe from digestion and blocks the downstream reaction to keep a highly fluorescent state. To testify this rationale, we utilized nuclear factor-kappa B p50 (NF-κB p50) as a model TFs. Owing to the amplification strategy, this method exhibited high sensitivity toward NF-κB p50 with a limit of detection of 10 pM, and a broad linear range from 30 pM to 1.5 nM. Furthermore, this method could detect multiple TFs in human colon cancer DLD-1 cells and reflect the variation in their cellular levels after stimulation. Finally, by conducting an inhibition assay we revealed the potential of this method for screening TFs-targeted drugs and calculating the IC50 of corresponding inhibitors.
研究领域:
生物化学
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分子生物学
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生物技术
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癌症研究
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药物发现
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DOI
PubMed
高影响因子
ANALYTICAL BIOCHEMISTRY
·
2022
·
IF 3.191
Ping Wang,Yaoyao Xie,Huikai Ma,Jiancheng Liu,Chenxiang Liu,Wei Feng,Shoumin Xi
A novel ratiometric electrochemical aptasensor was proposed for carcinoembryonic antigen (CEA) detection based on exonuclease III (Exo III)-assisted recycling and rolling circle amplification (RCA) strategies. A thiolated ferrocene-labeled hairpin probe 2 (Fc-HP2) was fixed on the gold nanoparticles (AuNPs)-modified gold electrode (AuE) surface through Au–S bonds. The presence of CEA led to the release of trigger, which hybridized with the 3′-protruding of hairpin probe 1 (HP1) and triggered the Exo III cleavage reaction, accompanied by the releasing of trigger and generation of new DNA fragment which was used for the successive hybridization with Fc-HP2. After the Exo III cleavage process, the remaining Fc-HP2 fragments hybridized as primers with the RCA template to initiate the RCA process, and long single-stranded polynucleotides were produced for methylene blue (MB) binding. Such changes resulted in the signal of Fc (I Fc ) decreased and that of MB (I MB ) increased, achieving a linear relationship between I MB /I Fc and logarithm of CEA concentrations ranging from 1.0 pg mL −1 to 100.0 ng mL −1 with a detection limit of 0.59 pg mL −1 . Additionally, the developed aptasensor had been successfully applied to detect CEA in human serum samples. Therefore, the proposed strategy might provide a new platform for clinical detections of CEA.
研究领域:
生物化学
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分析化学
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生物医学工程
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DOI
PubMed
高影响因子
SENSORS AND ACTUATORS B-CHEMICAL
·
2019
·
IF 6.393
Rufeng Zhang,Yu Wang,Xiaonan Qu,Shasha Li,Yihan Zhao,Su Liu,Jiadong Huang
Herein, an Exonuclease III (Exo III)-powered DNA walking machine has been integrated into electrochemical sensing for label-free and highly sensitive detection of antibiotics. The electrochemical sensing interface is prepared by co-assembly of hemin aptamer-containing DNA track (DT) and DNA walker (DW)-locking probe (LP) duplex on gold electrode surface. The specific target and aptamer interaction triggers Exo III-assisted cyclic amplification, releasing the unlocking probe (UP) that is used for the deblocking of DW. Then the autonomous moving of DW on electrode surface is propelled via Exo III digestion of hybridized DT. The consumption of DT induces the formation of G-quadruplex/hemin DNAzyme that can catalyze the reduction of H 2 O 2 , thus a significantly strong current signal is released. Using ampicillin as a model analyte, the proposed biosensor achieves a detection limit of 0.76 pM within a linear detection range from 1 pM to 10 nM. Additionally, this biosensing strategy can be readily extended for the detection of wide variety of analytes by using corresponding target recognition probes. Therefore, the proposed strategy indeed creates a highly sensitive and label-free electrochemical sensing platform for molecular diagnosis applications.
研究领域:
生物化学
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分析化学
·
分子诊断
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DOI
高影响因子
MICROCHIMICA ACTA
·
2019
·Vol. 186
·期 11
·1-9
·
IF 5.479
Liu Chao,Zhao Li,Liang Dongxia,Zhang Xiaoru,Song Weiling
A photocathode is described for the determination of microRNA-21 by using CuInS 2 as an active photocathode material. Exonuclease III assisted target recycling amplification was employed to enhance the detection sensitivity. The TATA-binding protein (TBP) was applied to enhance steric hindrance which decreases the photoelectrochemical intensity. This strategy is designed by combining the anti-interference photocathode material, enzyme assisted target recycling amplification and TBP induced signal off, showing remarkable amplification efficiency. Under the optimized conditions, the detection limit for microRNA-21 is as low as 0.47 fM, and a linear range was got from 1.0 × 10 −15 M to 1.0 × 10 −6 M.
Schematic representation of sensitive photoelectrochemical detection of microRNA-21.CuInS 2 is used as an active photocathode material. Combined Exonuclease III assisted target recycling amplification and TATA-binding protein decreased of photoelectrochemical intensity, the detection limit was 0.47 fM with good selectivity. (miR-21: microRNA-21; CS: chitosan).
研究领域:
生物化学
·
分析化学
·
分子生物学
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PubMed
高影响因子
SENSORS AND ACTUATORS B-CHEMICAL
·
2017
·
IF 5.401
Lili Yu,Wei Lan,Hui Xu,Hou Chen,Liangjiu Bai,Wenxiang Wang
We constructed a fluorometric and sensitive analysis of mercury (II) based on toehold-mediated strand displacement reaction, exonuclease III (Exo III) assisted target recycling, hybridization chain reaction (HCR), fluorescence intercalator SYBR Green I (SG) and grapheme oxide (GO). Firstly, toehold-mediated strand displacement reaction was constructed by using thymine-Hg 2+ -thymine (T-Hg 2+ -T) recognition mechanism between the helper hairpin DNA and assisted DNA. And then, the Exo III disintegrated the double-stranded DNA (dsDNA) by catalyzing the gradual removal of mononucleotides from 3′-hydroxyl terminus of dsDNA with a blunt 3′ terminus. It resulted in the target and assisted DNA recycling and released a trigger strand DNA at the same time. Finally, the trigger strand DNA initiated the hybridization chain reaction (HCR), forming long double helices, which achieved a secondary amplification due to the continuous target recycling, trigger strand DNA release and HCR. The number of T-T mismatches, the concentration of SG, Exo III and GO, the incubation time of Exo III were all optimized. Under the optimized condition, the limit of detection for Hg 2+ can be down to 7.37 pM with a linear range from 0 to 1.5 nM. This sensor exhibits high selectivity against other metal ions, and it works well for real samples.
研究领域:
分析化学
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纳米技术
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环境科学
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生物技术
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DOI
高影响因子
FOOD CONTROL
·
2022
·
IF 6.652
Chengyuan Wu,Xiangchuan Wu,Fan Hou,Lina Wu,Guangxia Liu
An ultrasensitive electrochemical aptasensor was established based on metalloporphyrin metal organic frameworks ( [email protected] ) with peroxidase-like activity and exonuclease III-assisted recycling magnification for the detection of ochratoxin A (OTA). [email protected] showed well peroxidase-like activities due to Fe-tetrakis (4-carboxyphenyl) porphyrin (Fe-TCPP) as a heme-like ligand and in situ supported with palladium nanoparticle, which could be used for the label of ssDNA instead of nature enzyme. The designed exonuclease III-assisted recycling magnification further improve the sensitivity for the detection. Based on this, the proposed aptasensor showed a well linear relationship within the scope of 1 × 10 −5 ng mL −1 to 10 ng mL −1 and a low detection limit of 6.79 fg mL −1 . Additionally, the application in the real spiked oil samples confirmed the dependability and veracity of the aptasensor. Hence, the designed aptasensor has comprehensive application prospects for the detection of OTA in food .
研究领域:
食品安全
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分析化学
·
生物技术
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产品关联
DOI
高影响因子
MICROCHEMICAL JOURNAL
·
2025
·
IF 5.1
Dianbo Yu,WenZhao Zhang,Jiahou Xu,Yuanxun Gong,Jiayi Zhang,Jihua Wei,Kai Zhang,Xianjiu Liao
Nuclear factor kappa B (NF-κB) p50 is a critical transcription factor that regulates immune responses, inflammation, and cell proliferation. Precise detection of NF-κB p50 at ultralow concentrations is of paramount importance for early disease diagnosis and mechanistic studies. Here, we report an electrochemical biosensor integrating double-crossover DNA tiles and entropy-driven strand displacement, amplified by Exonuclease III (Exo III) digestion. By designing a self-assembled DNA tile scaffold on a gold electrode and employing a DNA tetrahedron architecture, the sensor achieves a highly ordered surface configuration, minimizing background noise and enhancing specificity. In the absence of NF-κB p50, Exo III selectively digests a double-stranded DNA substrate, releasing single-stranded triggers that undergo an entropy-driven reaction to form a Y-shaped scaffold on the electrode. This assembly effectively hinders redox probe access, lowering the differential pulse voltammetry (DPV) signal. Conversely, NF-κB p50 binding protects the DNA substrate from digestion, preserving a higher current output. Under optimized conditions—30 nM DNA tiles, 2 U μL −1 Exo III, 30 min cleavage, and 100 nM DNA3/DNA4/DNA5—the biosensor exhibits an ultralow detection limit of 2.65 aM and excellent specificity, as confirmed by negligible cross-reactivity with other proteins. Furthermore, the method shows robust anti-interference performance in complex matrices (e.g., diluted cell lysates). These findings underscore the potential of this DNA tile-based biosensor for clinical diagnostics and fundamental research on NF-κB p50–mediated pathways.
研究领域:
生物传感器
·
分子诊断
·
生物电子学
·
核酸纳米技术
·
转录因子
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DOI
高影响因子
ANALYTICA CHIMICA ACTA
·
2020
·
IF 5.977
Na Wu,Yi-Ting Wang,Xiao-Yan Wang,Xu- Wei Chen,Ting Yang,Jian-Hua Wang
Rapid, sensitive, and user-friendly nucleic acid detection is of growing importance in early clinical diagnosis. Here, we construct a simple, one-pot and ultrasensitive DNA sensor via exonuclease III (Exo III)-assisted target recycling amplification (ERA) combined with 3D DNA walker cascade amplification. In the presence of single-stranded DNA target, the ERA process is activated to generate numerous walker strands (WS). Thereafter, Exo III-powered WSs autonomously move along magnetic bead (MB)-based 3D track to release numerous AgNCs into the supernatant as an amplified signal output. This biosensor had a low detection limit of 18 fM and an analytical range of 40 fM to 1 pM. Furthermore, the practical application potential of this biosensor was also confirmed by the spiking experiments of p53 into human serum and urine samples.
研究领域:
生物技术
·
分子生物学
·
分析化学
·
临床诊断
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产品关联
DOI
PubMed
高影响因子
危险品化学品经营许可证（带存储）