JOURNAL OF AGRICULTURAL AND FOOD CHEMISTRY
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2021
·
IF 5.279
Yiming Xie,Haiyan Wang,Xinyue Yuwen,Guosong Lai
Herein, we combine the exonuclease III (Exo III)-catalyzed release of a Zn2+-dependent ligation DNAzyme with the DNAzyme-driven strand displacement reaction (SDR) to develop a novel homogeneous colorimetric bioassay method for kanamycin (Kana) antibiotic detection. Upon the biorecognition reaction between Kana and a designed hairpin DNA, the DNAzyme-containing strand can be catalytically released by Exo III. Then, this DNAzyme will catalyze the ligation of two oligonucleotides to cause a SDR and the aggregation of gold nanoparticles (Au NPs) labeled by two linker DNA strands. Due to the aggregation of Au NPs for colorimetric signal transduction and the Exo III and SDR-assisted dual signal amplification, this method shows a wide linear range of 5 orders of magnitude and a very low detection limit down to 8.1 fg mL–1. Together with its excellent selectivity, repeatability, reliability, and convenient manipulation, the proposed method shows a great potential for the food quality monitoring application.
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MICROCHIMICA ACTA
·
2015
·Vol. 182
·期 13
·2251-2258
·
IF 3.741
Wang Minghua,Zhang Shuai,Ye Zihan,Peng Donglai,He Linghao,Yan Fufeng,Yang Yanqin,Zhang Hongzhong,Zhang Zhihong
Multilayered reduced graphene oxide (rGO) was functionalized with amino groups by treatment with nitrogen plasma. Raman spectroscopy showed plasma treatment not to substantially alter the chemical structure of rGO and that a wide range of functional nitrogen groups is evenly incorporated into the carbon lattice. The amino-modified rGO was used to design an electrochemical biosensor in which a DNAzyme, substrate DNA and Pb(II) and Hg(II) binding DNA were immobilized on the amino-rGO placed on a gold electrode. The high concentration of amino groups and the rough surface of the rGO favor DNA immobilization. Heavy metal ions are bound to the surface via specific interaction between DNA and the two ions which are detected by electrochemical impedance spectroscopy at a potential of 0.2 V (vs. Ag/AgCl). The detection limits for Pb(II) and Hg(II) are as low as 7.8 and 5.4 pM, respectively, and the analytical ranges extend from 0.01 to 100 nM. The sensor is highly specific and stable and therefore represents a highly promising tool for use in environmental monitoring.
A nanofilm of reduced graphene oxide was first modified with amino groups by treatment with nitrogen plasma. A special DNA was then anchored to the surface to obtain a biosensor for simultaneous detection of Pb(II) and Hg(II). The sensor has detection limits as low as 7.8 and 5.4 pM and is highly selective.
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MICROCHIMICA ACTA
·
2017
·Vol. 184
·期 6
·1791-1799
·
IF 4.58
Wang Xiangyan,Sun Duanping,Tong Yanli,Zhong Yongsheng,Chen Zuanguang
A regenerable and ultrasensitive voltammetric biosensor is described for the determination of thrombin. It relies on a combination of (a) enzymatic catalysis, (b) a G-quadruplex/hemin DNAzyme system, and (c) AuPd nanoparticles for signal amplification. Poly(o-phenylenediamine) was decorated with AuPd nanoparticles and loaded with horseradish peroxidase (HRP) and thrombin aptamer (TBA), and the mixture was allowed to interact with hemin to form the G-quadruplex/hemin/HRP/AuPd/poly(o-phenylenediamine) bioconjugates. In the presence of thrombin, the bioconjugates remain immobilized on the surface of the modified glassy carbon electrode through a sandwich reaction. Poly(o-phenylenediamine) also acts as a redox mediator, and the electrochemical reaction of poly(o-phenylenediamine) in the presence of H 2 O 2 is efficiently catalyzed by HRP, AuPd nanoparticles and G-quadruplex/hemin as the peroxidase mimics. Thus, a remarkably amplified electrochemical signal is obtained by the triple catalytic amplification. The biosensor has a dynamic range that spans the 100 f. to 20 nM thrombin concentration range, and the detection limit is 20 fM. The biosensor can be regenerated by applying an electrochemical desorption technique that breaks the gold-thiol bond and releases the components from the surface. In our perception, the mediator-free and signal-amplified biosensor demonstrated here has a large potential with respect to the quantitation of thrombin in clinical samples.
Schematic of the assay: The peroxidase-like AuPd nanoparticles, G-quadruplex/hemin DNAzyme and horseradish peroxidase synergistically catalyze the electrochemical reaction of poly(o-phenylenediamine) in the presence of H 2 O 2 , thereby amplifying the electrochemical detection signal of thrombin.
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CHEMICAL ENGINEERING JOURNAL
·
2024
·
IF 13.3
Zehan Zeng,Mingzhen Zhong,Tong Liao,Xuan Zhou,Lin Shi,Xin Zhang,Ye Zhang,Jingyan Feng,Yiqian Chen,Shuya Sheng,Jun Chen,Lei Zheng,Weilun Pan,Jinxiang Chen
Plasma extracellular vesicles (EVs) have emerged as promising biomarkers for the early diagnosis of glioblastoma. However, rapid and efficient isolation and detection of tumor-specific EV subsets from plasma remains challenging. To address this, we developed an innovative integrated chip based on self-sacrificial metal–organic frameworks and the epidermal growth factor receptor (EGFR) aptamers called ZIF-8@EGFR aptamer (ZIF-8@Apt), which further coupled with a “nano-fuels”-driven DNAzyme amplification strategy. The core functionality of the constructed chip lies in its ability to capture EVs with the ZIF-8@Apt module. Once captured, numerous DNA “nano-fuel” structures are in situ assembled on the EVs’ surfaces. Subsequently, the ZIF-8@Apt@EV@nano-fuels composites are lysed to release Zn 2+ , the EVs’ nucleic acids, and additional nano-fuels for subsequent detection. Finally, a DNAzyme detection system is introduced, where the released nano-fuels activate the DNAzyme, leading to multiple signal amplification aided by Zn 2+ for highly sensitive detection of plasma EVs. This integrated approach effectively enriches and detects target EVs, achieving a significantly enhanced detection sensitivity of 500 particles/μL. In a clinical study involving glioblastoma patients, our chip demonstrated a remarkable ability to distinguish between cancer patients and healthy individuals, achieving an area under the curve (AUC) of 0.958. Moreover, the response signals of the chip can be conveniently read using fluorescence spectroscopy or microscope, offering a rapid, highly sensitive, and high-throughput analysis of plasma EVs for various application scenarios. Overall, this study introduces a comprehensive approach for the rapid isolation and accurate analysis of plasma EVs, holding significant promise for early diagnosis and disease monitoring.
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Advanced Science
·
2024
·
IF 14.3
Huiyan Du,Ensheng Xu,Yihan Xu,Qingwang Xue,Hongxia Xu,Jibin Song
The development of theragnostic nanosystems integrating FRET (fluorescence resonance energy transfer) imaging and chemodynamic therapy (CDT) for accurate diagnosis and effective treatment of lung tumors is still a big challenge. Herein, a peptide-assembled 3D DNAzyme motor nanodevice is engineered for a self-powered FRET amplifier profiling human neutrophil elastase (HNE) and self-supplied H 2 O 2 enhancing CDT. The nanodevice is prepared by depositing AuNPs on ZIF-8, in which ZIF-8 co-loaded the lysosomal targeting peptide-modified copper peroxides (PCPs) and hairpins (H1, H2, and H3), AuNPs are co-labeled by DNAzyme-peptide (DP) conjugate and H3. In the tumor micro-environment, HNE driven 3D DNAzyme walker followed by an exponential amplification constructed by a synergistic cross-activation between hybridization chain reaction and DNAzyme, generating a self-powered FRET amplifier. The FRET amplifier specifically measures HNE with a sensitivity of 0.026 pM, and successfully images exogenous HNE in living cells and monitors HNE in mouse models. Moreover, the PCPs can target lysosomes, reducing lysosome escape. The self-supplying H 2 O 2 undertaken by PCPs improves the Cu (II)-catalyzed Fenton-like reaction, effectively causing cell apoptosis to inhibit tumor growth. Significantly, the nanodevice successfully screens inhibitors and discriminates the HNE level in normal and lung cancer tissues, suggesting that the nanodevice provides an effective tool for the diagnosis and treatment of lung tumors.
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SCIENCE OF THE TOTAL ENVIRONMENT
·
2021
·
IF 7.963
Nuanfei Zhu,Xuesong Li,Ye Liu,Jingfu Liu,Yawei Wang,Xiangyang Wu,Zhen Zhang
Dibutyl phthalate (DBP) is considered as one of the most widely used phthalate esters (PAEs), which has attracted worldwide concerns because of its potential threats to eco-environments and human health. Systematic investigations of DBP environmental occurrence contribute to the further risk assessment, which depends on effective and available analytical methods. In this study, an amplified ratiometric fluorescence ELISA was established for sensitive and high-throughput detection of DBP in the aquatic system based on a novel tetrahedral DNA nanostructure (TDN)-scaffolded-DNAzyme (Tetrazyme). Wherein, Tetrazyme was prepared by the precise folding of G-quadruplex sequence on three vertex angles of the TDN, together with hemin as the horseradish peroxidase (HRP)-mimicking enzyme. The rigid TDN avoided the local overcrowding effect to provide a reasonable spatial spacing on the interface for G-quadruplex sequence, increasing the collision chance between DNAzyme and substrates, improving the catalytic ability of DNAzyme effectively. Besides, streptavidin (SA) and biotin (bio) were used to anchor TDN and antibody, in which the specific binding of SA/bio could make more Tetrazyme conjugate on each signal element, resulting in the dual signal amplification. Meanwhile, the accuracy and precision were enhanced owing to the inherent built-in rectification to the environment from the dual output ratiometric fluorescence assay. Under the optimized conditions, the detection limit of this proposed method was 0.17 ng/mL (16 times lower than that of conventional ELISA using the same antibody) with a satisfactory accuracy (recoveries, 79.0%- 116.2%; CV, 2.1–6.5%). Overall, this platform provides a promising way for accurate, sensitive and rapid determination of DBP from environmental waters.
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MICROCHIMICA ACTA
·
2019
·Vol. 186
·期 8
·1-8
·
IF 5.479
Song Xiaolei,Wang Yu,Liu Su,Zhang Xue,Wang Jingfeng,Wang Haiwang,Zhang Fenfen,Yu Jinghua,Huang Jiadong
An amplified electrochemical biosensing scheme is described for lead(II) ions. It is making use of DNAzyme-assisted target recycling and catalytic hairpin assembly (CHA). The hairpin strand (substrate probe for the Pb 2+ -based DNAzyme; referred to as SP) is composed of trigger probe (TP) and a capture probe 1 attached to gold nanoparticles (AuNP). In the presence of the enzyme probe that partially hybridizes with SP, the introduction of Pb 2+ triggers target recycling and drives the highly amplified translation of target Pb(II) to TP. The CHA reaction is further initiated by TP. The modified AuNP act as the connecting unit, and this leads to the formation of a 3D DNA-AuNP network on the electrode (which is the third amplification step). It can bind the positively charged redox mediator RuHex via electrostatic interaction for electrochemical detection. This biosensor has a low detection limit (95 pM) and any analytical range that covers the 100 pM to 5 μM Pb(II) concentration range. It is selective over other divalent metal ions. It was applied to the determination of Pb 2+ in spiked real-world samples.
Schematic presentation of the electrochemical biosensor. The triply amplified electrochemical assay is based on the use of DNAzyme-assisted target recycling with catalytic hairpin assembly (CHA) reaction for sensitive and selective determination of lead ion (Pb 2+ ). AuNP: gold nanoparticles; SP: substrate probe; EP: enzyme probe.
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MICROCHIMICA ACTA
·
2018
·Vol. 185
·期 9
·1-9
·
IF 5.705
Wang Jing,Li Haigang,Li Tingting,Ling Liansheng
A one-step, one-tube colorimetric assay is described for the detection of bacterial double-stranded DNA (dsDNA). It utilizes a G-quadruplex DNAzyme produced by an asymmetric polymerase chain reaction (As-PCR) that catalyzes the oxidation of cysteine to form cystine. This results in the formation of oligonucleotide-modified gold nanoparticles via triplex formation, and eventually in a color change from red to blue that occurs within about 10 mins. This can be measured by ratiometric colorimetric (at 525 and 600 nm). The limit of detection (LOD) for the model analyte (dsDNA of Staphylococcus aureus ( S. aureus )) is as low as 0.28 pg per 0.05 mL with a good linear response ranging from 16.0 fg·μL −1 to 1.6 ng·μL −1 . This is much lower than previously reported LODs. The assay is highly selective for S. aureus dsDNA over a range of other bacterial DNAs. Conceivably, it provides an attractive alternative tool for rapid detection of bacterial dsDNA as required in pathogen screening in the food industry.
Schematic presentation of a colorimetric assay for bacterial DNA. It is based on the catalytic activity of a G-quadruplex DNAzyme that is formed by an asymmetric PCR involving triplex DNA formation and gold nanoparticle (AuNPs) aggregation.
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TALANTA
·
2018
·
IF 4.244
Qian Luan,Xin Xiong,Ning Gan,Yuting Cao,Tianhua Li,Dazhen Wu,Youren Dong,Futao Hu
An ultrasensitive colorimetric aptasensor was developed for antibiotics detection, with chloramphenicol (CAP) as model target, using DNAzyme labeled Fe-MIL-88-Pt as novel peroxidase mimic signal tags and target-triggered circular strand-displacement polymerization (CSDP) for signal amplification. The system consists of two components which can partially hybridize with each other: one is capture probe which was formed through immobilizing hairpin DNA containing aptamer sequence on magnetic beads (MB-cDNA), another is signal tag which was constructed through labeling single strand DNAzyme (G-quadruplex/Hemin) which can partially hybrid with cDNA on platinum nanoparticles functionalized Fe-MIL-88 (MIL-88-Pt-DNAzyme). All components of MIL-88, Pt and DNAzyme in the tag can act as peroxidase mimic to triply catalyze the oxidation of 3,3′,5,5′-tetramethylbenzidine (TMB) by H 2 O 2 into a blue-colored oxidized TMB (oxTMB) for the colorimetric readout. Thus distinctive signal can be observed by naked eye even in presence of 0.02 nM tags. In the presence of target and primer, cDNA loop can open to form cDNA/CAP intermediates, enabling primer to hybridize with the exposed sequences of the cDNA, which initiated target assisted CSDP recycles. Then numerous signal tags were released into supernatant to catalyze TMB for color development. There was a liner relationship between the absorbance and the concentration of CAP in the range of 0.1 pM (0.0323 pg/mL) to 1000 pM (323 pg/mL) with the detection limit of 0.03 pM (0.0097 pg/mL). The ultra-high sensitivity was ascribed to the multiplex catalytic activities from the tags and CSDP based signal amplification. Furthermore, this method can produce signals being observed by naked eye to facilitate in-situ detection and be further extended to detect other antibiotics in food just by simply replacing cDNA on the sensing system.
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Food Analytical Methods
·
2014
·Vol. 8
·期 6
·1556-1564
·
IF 1.802
Liu Tao,Zhang Xiao,Hao Jianpeng,Zhu Wenxin,Liu Wei,Zhang Daohong,Wang Jianlong
Herein, we for the first time report an acetylcholinesterase (AChE)-free colorimetric method for the detection of chlorpyrifos (CP), which is based on the oxidation reaction of H 2 O 2 with CP and 2,2′-Azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) diammonium salt (ABTS 2− ) catalyzed by hemin/G-quadruplex DNAzyme. In the absence of CP, ABTS 2− is directly oxidized by H 2 O 2 with hemin/G-quadruplex DNAzyme as a biocatalyst to produce the blue-green-colored free radical anion (ABTS ·− ), accompanied with an obvious UV-visible absorbance at 418 nm. However, the amount of H 2 O 2 used in H 2 O 2 -DNAzyme-ABTS 2− system will be decreased if CP is first reacted with H 2 O 2 , leading to the decrease of the amount of ABTS ·− and a change in the UV-visible absorbance of ABTS ·− at 418 nm. Thus, an indirect dependence relation between CP concentration and the absorbance of ABTS ·− at 418 nm could be constructed and used for CP detection. Under optimal conditions, the detection range of the method is 0.04–1 μg/mL, with a detection limit of 0.01 μg/mL. Application of the method in real juice samples shows good analytical features in terms of recoveries (95–107.5 %), the relative standard deviation (RSD) (0.6–4.7 %), rendering it as an attractive candidate to current methodologies for the determination of CP.
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MICROCHEMICAL JOURNAL
·
2026
·
IF 5.2
Beili Wang,Wei Gao,Liang Yang
An Exo-III-triggered target-reconstructed DNAzyme walker enables ultrasensitive multiplex miRNA detection. • Cascade amplification integrates Exo-III recycling, polymerase/endonuclease cycling, and DNAzyme walking. • Orthogonal Pt, Ir, Rh tags allow simultaneous ICP-MS quantification of three miRNAs without spectral cross-talk. • Detection limits reach 0.15 fM (miR-21), 0.32 fM (miR-1246), and 0.26 fM (miR-20a) with excellent linearity. • The method discriminates single-base mismatches and performs well in diluted human serum (92–108% recovery). Cervical cancer remains a leading cause of cancer-related mortality among women, and the prognosis of patients after radiotherapy varies considerably due to differences in tumor radiosensitivity and treatment response. Reliable biomarkers for post-radiotherapy prognosis are therefore critically needed. MicroRNAs (miRNAs) have emerged as promising non-invasive indicators, yet accurate detection is hindered by their low abundance and the lack of multiplexing capability in conventional methods. Herein, we report a highly sensitive and specific platform for simultaneous detection of three cervical cancer radiotherapy-associated miRNAs (miR-21, miR-1246, and miR-20a) by integrating an Exo-III-triggered target-reconstructed DNAzyme walker with inductively coupled plasma mass spectrometry (ICP-MS). The strategy employs a structurally locked DNA walker that is sequentially activated through Exo-III-mediated target recycling and DNA polymerase/endonuclease-assisted cyclic amplification, ultimately unleashing a DNAzyme walker to cleave element-labeled substrates. Using orthogonal platinum group element tags (Pt, Ir, Rh), the released tags are quantified by ICP-MS without spectral cross-talk. The method achieves detection limits as low as 0.15 fM, 0.32 fM, and 0.26 fM for miR-21, miR-1246, and miR-20a, respectively, with excellent single-nucleotide discrimination and negligible cross-reactivity. Successful spike-recovery experiments in diluted human serum validate its practical feasibility. This cascade amplification strategy offers a robust, multiplex, and ultrasensitive tool for non-invasive liquid biopsy-based prognostic assessment of post-radiotherapy cervical cancer, and can be readily adapted for other disease-related miRNA panels. Download: Download high-res image (121KB) Download: Download full-size image
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Analytical Methods
·
2026
·
IF 3
Yi Tian,Jiaxuan Chen,Fengzheng Chen,Yu Ling,Shengyue Zheng,Daizhen Jian,Qiong Tang,Song Wang,Kai Shi
Split crRNA with Cas12a (SCas12a)-powered biosensor has shown sensitivity and accuracy in nucleic acid detection, but its application to metal ion target detection still needs to be explored. By integrating an SCas12a-mediated cascade strand displacement reaction (CSDR) with a phosphorothioate-modified hairpin G-quadruplex (psHG4) reporter, a portable “turn-on” colorimetric/photothermal system was engineered for the precise and sensitive detection of Hg2+ (termed as SCas12a/CSDR-psHG4). Target Hg2+ triggers SCas12a-mediated CSDR, and then activates SCas12a’s nuclease activity. Activated SCas12a cleaves the loop region of psHG4, thereby liberating the previously silenced psG4 sequence, which subsequently assembles into a psG4/hemin complex in the presence of K⁺ and hemin. This complex catalyzes the typical oxidation of 3,3′,5,5′-tetramethylbenzidine (TMB), generating the colorimetric and photothermal product TMBox. Record the RGB value of the solution with a portable smartphone and track the temperature change using a thermometer. This colorimetric/photothermal biosensor displayed a linear range of 1 pmol/L to 100 nmol/L and 100 fmol/L to 10 nmol/L, with a detection limit of 0.5 pmol/L and 60 fmol/L, respectively. Utilizing the SCas12a technique, the SCas12a/CSDR-psHG4 presents a fresh method for identifying metal ion targets, along with significant prospects for enhancing human health protection and environmental surveillance.
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