JOURNAL OF ELECTROANALYTICAL CHEMISTRY
·
2020
·
IF 3.807
Xiao Yang,Minghui Feng,Jianfei Xia,Feifei Zhang,Zonghua Wang
A novel electrochemical biosensor based on AuNPs/Ti 3 C 2 MXene three-dimensional nanocomposite for sensitive miRNA-155 detection by exonuclease III (Exo III)-aided cascade target recycling is reported. Ti 3 C 2 MXene is synthesized and used as a support to immobilize Au nanoparticles (AuNPs) due to its special layered structure and metallic properties. The three-dimensional (3D) structure of the AuNPs/Ti 3 C 2 MXene nanocomposite for the biosensor platform leverages the integrated advantages of a large specific surface area, excellent electrical conductivity and electrocatalytic properties . AuNPs are used to immobilize capture DNA (C-DNA) by Au S chemical bonds. The C-DNA was modified with methylene blue (MB) at the 3′ end, resulting in an initial DPV electrochemical signal ( I d ). In the presence of target, miRNA-155 and C-DNA formed a double-stranded structure by complementary base pairing. Subsequently, Exo III digested the 3′ end of the C-DNA in the double-stranded structure and caused the electrochemical signal to “turn off” (the final signal denoted as I h ). By Exo III-aided cascade cleavage of the target, one miRNA-155 chain causes the release of several C-DNAs under certain conditions, which significantly amplifies the peak current difference on the fabricated biosensor (Δ I = I d ﹣ I h ). The electrochemical sensor achieved a detection limit of 0.35 fM (S/N = 3) with a linear range from 1.0 fM to 10 nM. This sensor also shows desirable stability, reproducibility and specificity.
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高影响因子
MICROCHIMICA ACTA
·
2021
·Vol. 188
·期 8
·1-9
·
IF 5.833
Yang Jiao,Fu Shilan,Luo Fang,Guo Longhua,Qiu Bin,Lin Zhenyu
MicroRNAs can serve as biomarkers for many cancers, so it is significant to develop simple and sensitive strategies for microRNAs detection. Photoelectrochemical (PEC) detection has the advantages of simple equipment and high sensitivity. But in conventional PEC DNA sensors, tedious immobilization procedures of photoactive materials and capture probes on electrode surfaces are inevitable. To overcome those limitations, a homogeneous PEC biosensor based on target-responsive hydrogels has been developed (miRNA-155 has been chosen as a model target). PEC signal molecules (TiO 2 nanoparticles, TiO 2 NPs) were embedded in DNA hydrogels formed by hyaluronic acid sodium salt, amine-modified DNA double strands, and polyethylenimine rich in amine groups. In the presence of the target, DNA double strands in hydrogel were nicked by endonuclease and TiO 2 NPs were released to the supernate and a high PEC response was obtained when collecting the supernate for PEC test, while almost no TiO 2 NPs released in the absence of the target. Thanks to the exonuclease III and nicking endonuclease Nb.BbvCI-assisted cascaded amplification strategy, the proposed biosensor exhibits high sensitivity toward miRNA-155 with a low detection limit of 0.41 fM and a wide linear range from 1.0 fM to 100 pM. Since this method circumvents tedious electrode modification procedures, the proposed technique exhibits the advantages of simplicity and good reproducibility. Moreover, the prepared hydrogels have outstanding storage stability, so that they can be prepared in advance and shorten detection time. This biosensing platform provides a versatile strategy for the construction of homogeneous PEC biosensors for the detection of diverse targets. Graphical abstract
Photoelectrochemical detection techniques have been coupled with controlled release system to develop an immobilization-free microRNA biosensor. High sensitivity has been realized based on cascaded signal amplification strategy, and the proposed biosensor has been applied to detect the target in real sample with satisfied results. Since no tedious electrode modifications, the proposed homogeneous PEC sensor exhibits high reproducibility and good stability.
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高影响因子
SENSORS AND ACTUATORS B-CHEMICAL
·
2019
·
IF 6.393
Yu Ling,Jiao Zhou,Xiao Fang Zhang,Xiao Hu Wang,Nian Bing Li,Hong Qun Luo
As one of critical base excision repair enzymes, uracil-DNA glycosylase (UDG) can specifically repair uracil-induced DNA damage to maintain the genome integrity of organisms. Moreover, abnormal expression of UDG is related to various cancers and other serious diseases. Therefore, it is essential to accurately and sensitively monitor UDG activity. In this work, a ratiometric fluorescence method using poly-(thymine) DNA template copper nanoclusters (Cu NCs) and 4′,6-diamidino-2-phenylindole (DAPI) as the output signals is developed for simple, selective, and sensitive detection of UDG. A double-stranded DNA (dsDNA) consisting of a uracil-containing single chain and a trigger sequence is designed as a substrate dsDNA. Meanwhile, another dsDNA comprising 30 pairs of A-T bases ((AT) 30 dsDNA) is designed to be partially complementary to the trigger chain. The (AT) 30 dsDNA itself has no blunt or recessed 3′ end and cannot be cut by exonuclease III (Exo III). With the Exo III-assisted amplification, adding UDG results in the success synthesis of red-emissive Cu NCs, while the dsDNAs in the system are hydrolyzed and the fluorescence signal of DAPI is weak. In contrast, without addition of UDG, the fluorescence intensity of Cu NCs is low, while the DAPI emits significantly enhanced fluorescence signal. The proposed method presents a detection limit of 5.0 × 10 −5 U mL −1 and can be applied to the HeLa cell lysate sample with satisfactory results. Additionally, the kinetic parameter is studied and the UDG inhibition effect is evaluated. In summary, this work provides a potential platform for DNA repair enzyme-related analysis and clinical diagnosis.
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高影响因子
BIOSENSORS & BIOELECTRONICS
·
2015
·
IF 6.409
Sai Bi,Xiaoqiang Jia,Jiayan Ye,Ying Dong
Gold nanomaterials promise a wide range of potential applications in chemical and biological sensing, imaging, and catalysis. In this paper, we demonstrate a facile method for room-temperature synthesis of gold nanostars (AuNSs) with a size of ~50 nm via seeded growth. Significantly, the AuNSs are found to have high light-scattering properties, which are successfully used as labels for sensitive and selective detection of nucleic acids and proteins by using exonuclease III (Exo III) as a biocatalyst . For DNA detection, the binding of targets to the functionalized AuNS probes leads to the Exo III-stimulated cascade recycling amplification. As a result, a large amount of AuNSs are released from magnetic nanoparticles (MNPs) into solution, providing a greatly enhanced light-scattering signal for amplified sensing process. Moreover, a binding-induced DNA three-way junction (DNA TWJ) is introduced to thrombin detection, in which the binding of two aptamers to thrombin triggers assembly of the DNA motifs and initiates the subsequent DNA strand displacement reaction (SDR) and Exo III-assisted cascade recycling amplification. The detection limits of 89 fM and 5.6 pM are achieved for DNA and thrombin, respectively, which are comparable to or even exceed that of the reported isothermal amplification methods. It is noteworthy that based on the DNA TWJ strategy the sequences are independent on target proteins. Additionally, the employment of MNPs in the assays can not only simplify the operations but also improve the detection sensitivity. Therefore, the proposed amplified light-scattering assay with high sensitivity and selectivity , acceptable accuracy, and satisfactory versatility of analytes provides various applications in bioanalysis .
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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.
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BIOELECTROCHEMISTRY
·
2024
·
IF 4.8
Shan Huang,Yali Wang,Shuai Liu,Huihao Li,Mingli Yang,Yi Fang,Qi Xiao
Carcinoembryonic antigen (CEA), a key colon biomarker, demands a precise detection method for cancer diagnosis and prognosis. This study introduces a novel electrochemical aptasensor using a triblock polyadenine probe for ultra-sensitive detection of CEA. The method leverages Exonuclease III (Exo III)-assisted target recycling and hybridization chain reaction. The triblock polyadenine probe self-assembles on the bare gold electrode through the strong affinity between adenine and gold electrode, blocking CEA diffusion and providing a large immobilization surface. CEA binding to hairpin probe 1 (HP1), followed by the hybridization between HP1 and hairpin probe 2 (HP2), triggers DNA cleavage by Exo III, amplifying the signal via a hybridization chain reaction and producing numerous dsDNA walkers that generates a dramatic electrochemical impedance signal. Under optimized conditions, the aptasensor achieved two ultra-low detection limits: 0.39 ag∙mL −1 within the concentration range of 5 ag∙mL −1 to 5 × 10 6 ag∙mL −1 , and 1.5 ag∙mL −1 within the concentration range of 5 × 10 6 ag∙mL −1 to 1 × 10 10 ag∙mL −1 . Its performance in human serum samples meets the practical standards, offering a promising new tool for ultrasensitive tumor marker detection, potentially revolutionizing early cancer diagnosis.
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MICROCHEMICAL JOURNAL
·
2022
·
IF 5.304
Meijuan Du,Qian Chen,Xueqin Xu
A novel and label-free electrochemical aptasensor based on exonuclease III (Exo III) and G-quadruplex DNAzyme has been first designed to detect metronidazole (MNZ) sensitively and selectively in this paper. Capture probe (CP), helper probe (HP) and aptamer (Apt) hybridize to form double-stranded DNA (dsDNA) structure with a blunt 3′-terminus, resulting in the guanine (G)-rich CP being enclosed in the dsDNA, followed by specific digestion of the 3′-terminus by Exo III to expose the G-rich sequence. At this time, CP can bind with the added hemin to form G-quadruplex DNAzyme which catalyzes the redox reaction of 3,3′,5,5′-tetramethylbenzidine (TMB) and H 2 O 2 , generating a strong current signal. However, in the presence of MNZ, the binding of MNZ and aptamer breaks the original blunt 3′-terminus of the dsDNA structure, blocking the subsequent series of reactions, and only a weak current signal can be observed. Under the optimal conditions, the aptasensor showed excellent sensitivity and specificity for the detection of MNZ with a wide linear range from 10 pM to 500 nM and a detection limit as low as 3.0 pM (S/N = 3). The aptasensor constructed based on MNZ aptamer can effectively avoid the interference of other nitro compounds and antibiotics to a certain extent. Compared with other specific recognition elements, the aptasensor possesses the advantages of strong specificity, simple preparation and low cost. Moreover, the proposed aptasensor has been successfully applied to the detection of MNZ in tap water and honey samples.
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高影响因子
Journal of Materials Chemistry B
·
2015
·Vol. 3
·期 41
·8180-8187
·
IF 4.726
Ke-Jing Huang,Yu-Jie Liu,Qiu-Fen Zhai
Recently, two-dimensional layered transition metal dichalcogenide materials have been very fascinating in electrochemical fields due to their unique electronic and electrochemical properties. In the present work, a novel VS2–graphene (VS2–GR) composite is firstly prepared by a facile one-step hydrothermal method. A sensitive, universal and label-free electrochemical aptamer-based sensor (aptasensor) is then designed to detect the platelet-derived growth factor BB (PDGF-BB) based on VS2–GR coupled with Exo III-aided autocatalytic signal amplification. In the absence of PDGE-BB, the aptamer hybridized with the complementary DNA (cDNA) and the single-strand signal DNA labeled with biotin at 5′ end cannot be cleaved by the Exo III. When the above mixture is applied on the tetrahedron-structured probe (T-DNA) modified AuNP/VS2–GR electrode, signal DNA hybridizes with the protruding of T-DNA and a large number of avidin-HRP are adsorbed on the modified electrode by the specific binding of avidin–biotin, which leads to a strong current response by the catalysis of HRP to the mixture of hydrogen peroxide and hydroquinone (H2O2 + HQ). However, in the presence of PDGF-BB, the aptamer is preferentially associated with PDGF-BB, which facilitates the formation of double-strand DNA between cDNA and signal DNA. The duplex DNA is digested by Exo III from the 3′ blunt terminus of signal DNA and the cDNA is released. The liberative cDNA then reacts with the remaining signal DNA in the solution to perform a new cleavage process. Finally, the cyclic hybridization–hydrolysis process leads to a great decrease of biotin on the electrode, thus reducing the quantity of signal molecule HRP and causing an obvious decline of current response. This proposed protocol exhibits high specificity, good sensitivity, and good stability for PDGF-BB detection. Moreover, the aptasensor can be applied as a universal assay for the detection of various targets including proteins, DNAs, and metal ions.
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高影响因子
ANALYTICA CHIMICA ACTA
·
2026
·
IF 6.1
Xiaodong Cao,Mingli Liu,Yifei Pan,Qiuran Wang,Jinying Gong,Yongkang Ye,Yuhan Sun,Zhaojun Wei
Sensitive and simultaneous detection of multiple foodborne pathogens has become an urgent need in the food industry. This work presents a signal-on fluorescence biosensor using DNA-templated metal nanoclusters as signal nanotags and an Exonuclease III (Exo III)-assisted target-recycling (EATR) strategy for the detection of Cronobacter sakazakii ( C. sakazakii ) and Salmonella typhimurium ( S. typhimurium ). The detection system consists of carboxylated polypyrrole nanoparticles functionalized with hairpin DNA sequences HP1 and HP2 (HP1-cPPy-HP2 NPs), the target ompA gene (tDNA1) of C. sakazakii and invA gene (tDNA2) of S. typhimurium , DNA-templated gold and silver nanocluster nanotags (sDNA1-AuNCs and sDNA2-AgNCs), and Exo III. In the presence of the targets, HP1 and HP2 on cPPy surface are opened through hybridization, forming tDNA1-HP1-cPPy-HP2-tDNA2 complex. These opened HP1 and HP2 then hybridize with their respective sDNA1 (sDNA1-AuNCs) and sDNA2 (sDNA2-AgNCs), leading to the increased fluorescence intensity of both AuNCs and AgNCs in the detection system. Furthermore, with EATR amplification strategy, the targets can be released from the complex through cleavage by Exo III and proceed to the next cycle of HP1/HP2 opening. As a result, the fluorescence signals of AuNCs and AgNCs can be significantly increased. Under optimal conditions, the fluorescence biosensor shows wide linear ranges for C. sakazakii (1.64 ∼ 1.64×10 5 CFU mL −1 ) and S. typhimurium (2.56 ∼ 2.56×10 5 CFU mL −1 ), with LODs of 1.4 CFU mL −1 and 2.2 CFU mL −1 , respectively. The biosensor was used for the detection of C. sakazakii and S. typhimurium in real samples, with recovery ranges of 88.8% to 110.6% and 90.4% to 114.4%, respectively.
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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).
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高影响因子
BIOSENSORS & BIOELECTRONICS
·
2015
·
IF 6.409
Ke-Jing Huang,Hong-Lei Shuai,Ji-Zong Zhang
A highly sensitive and ultrasensitive electrochemical aptasensor for platelet-derived growth factor BB (PDGF-BB) detection is fabricated based on layered molybdenum selenide–graphene (MoSe 2 –Gr) composites and Exonuclease III (Exo III)-aided signal amplification . MoSe 2 –Gr is prepared by a simple hydrothermal method and used as a promising sensing platform. Exo III has a specifical exo-deoxyribonuclease activity for duplex DNAs in the direction from 3′ to 5′ terminus, however its activity is limited on the duplex DNAs with more than 4 mismatched terminal bases at 3′ ends. Herein, aptamer and complementary DNA (cDNA) sequences are designed with four thymine bases on 3′ ends. In the presence of target protein, the aptamer associates with it and facilitates the formation of duplex DNA between cDNA and signal DNA. The duplex DNA then is digested by Exo III and releases cDNA, which hybridizes with signal DNA to perform a new cleavage process. Nevertheless, in the absence of target protein, the aptamer hybridizes with cDNA will inhibit the Exo III-assisted nucleotides cleavage. The signal DNA then hybridizes with capture DNA on the electrode. Subsequently, horse radish peroxidase is fixed on electrode by avidin–biotin reaction and then catalyzes hydrogen peroxide and hydroquinone to produce electrochemical response. Therefore, a bridge can be established between the concentration of target protein and the degree of the attenuation of the obtained signal, providing a quantitative measure of target protein with a broad detection range of 0.0001–1 nM and a detection limit of 20 fM.
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高影响因子
SENSORS AND ACTUATORS B-CHEMICAL
·
2017
·
IF 5.401
Bing Zhang,Hongyun Meng,Xue Wang,Jing Li,Honghong Chang,Wenlong Wei
Herein we designed a novel photoelectrochemical sensing platform based on Fe 3+ doped ZnO-Ag photocatalyst (Fe 3+ /ZnO-Ag) for ultrasensitive Hg 2+ detection by using exonuclease III-assisted target recycling and DNAzyme-catalyzed amplification. Firstly, Fe 3+ /ZnO-Ag was synthesized to provide photocurrent under visible light irradiation. Secondly, target Hg 2+ was determined by hairpin DNA probes on the Fe 3+ /ZnO-Ag modified ITO based on T-Hg 2+ -T, and then Hg 2+ was released by the digestion of exonuclease III to double-stranded DNA. Finally, the photocurrent markedly decreased due to catalysis of hemin/G-quadruplex toward 4-chloro-1-naphthol. Under optimal conditions, the photocurrents were linearly related to Hg 2+ concentrations form 0. 5 nM to 100 nM with a detection limit of 0. 1 nM at the 3 S blank criterion. More importantly, the developed biosensor exhibited good selectivity, repeatability, and stability, meanwhile, this analysis platform held great potential for testing real water samples in future.
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