SENSORS AND ACTUATORS B-CHEMICAL
·
2015
·
IF 4.097
Weihua Zhu,Chenglei Xuan,Guoliang Liu,Zhuo Chen,Wei Wang
A simple fluorescent assay for sensing bovine serum albumin (BSA) and calf thymus DNA (ctDNA) was developed based on the gold nanorods-based probe. Gold nanorods coated with mesoporous silica coated (Au@SiO 2 ) which loading acridine orange (AO) was prepared by one-pot synthesis method. Interestingly, BSA is detected based on the quenching fluorescence signal while ctDNA is determined based on the enhanced fluorescence signal by the proposed method. Under the optimal conditions, the declined values of the fluorescent intensity of the biosensor are proportional to the BSA concentration in the range of 0.75–33.86 μmol/L. The limit of detection is 0.25 μmol/L. The enhanced values of the fluorescent intensity of the biosensor are linear with the ctDNA concentration in the range of 0.5–10 μg/mL, and the detection limit is 0.1667 μg/mL. The proposed method shows a good selectivity for BSA and ctDNA and can be applied to the determination of the analytes in real samples. The strategy of Au NRs@SiO 2 -AO complex as a novel fluorescent probe paves a new way to design fluorescent sensor.
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ANALYTICAL CHEMISTRY
·
2017
·
IF 6.32
Hao Hu,Jinyi Zhang,Yu Ding,Xinfeng Zhang,Kailai Xu,Xiandeng Hou,Peng Wu
Singlet oxygen (1O2), generated via photosensitization, has been proved to oxidize chromogenic substrates with neither H2O2 oxidation nor enzyme (horseradish peroxidase, HRP) catalysis. Of the various methods for modulation of the 1O2 generation, DNA-controlled photosensitization received great attention. Therefore, integration of the formation/deformation DNA structures with DNA-controlled photosensitization will be extremely appealing in visual biosensor developments. Here, the stable melamine-thymine complex was explored in combination with DNA-controlled photosensitization for visual detection of melamine. A T-rich single stand DNA was utilized as the recognition unit. Upon the formation of the T-M-T complex, double stand DNA was formed, which was ready for the binding of SYBR Green I and activated the photosensitization. Subsequent oxidation of TMB allowed visual detection of melamine in dairy products, with spike-recoveries ranging from 94% to 106%.
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Nanoscale
·
2017
·Vol. 9
·期 33
·11912-11919
·
IF 7.367
Siqi Zhang,Kun Wang,Kai-Bin Li,Fengzao Chen,Wei Shi,Wen-Ping Jia,Jie Zhang,De-Man Han
Molecular logic devices with different functions can perform various tasks in the areas of biological molecule detection, disease diagnosis, multivariate analysis, and bioimaging. Herein, a series of logic circuits based on silver nanoclusters (AgNCs)/graphene oxide (GO) are constructed to execute nonarithmetic functions, including 3-, 4-, and 5-bit odd/even checking. The resulting devices can differentiate between even and odd decimal numbers in the range from 0 to 31. Moreover, the devices can be expanded to operate with wider ranges of numbers when more inputs are added. The signal reporter is structured using AgNCs and GO, preventing laborious modification of biomolecules. The designed DNA-based logic nanodevices share the same DNA platform and a constant threshold value, showing great potential for application in information processing at the molecular level. Additionally, these devices can stably carry out their logic operations in a biological matrix, indicating that the AgNC/GO-based system can operate in a complicated biological environment. Given the biocompatibility and design flexibility of DNA, this study provides novel outcomes towards the development of label-free intelligent nanodevices. This may open a new path for the application of AgNCs/GO in molecular logic circuits and fluorescence imaging.
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JOURNAL OF NANOBIOTECHNOLOGY
·
2024
·Vol. 22
·期 1
·1-21
·
IF 10.6
Yu Jiang,Gao Yidan,Liu Fei,Zhang Yuyu,Li Jianda,Ding Luogang,Ren Sufang,Yang Jie,Jiao Jian,Feng Gong,Chen Zhi,Sun Wenbo,Wu Jiaqiang
Background Animal and human health are seriously threatened by bacterial infections, which can lead to bacteremia and extremely high rates of morbidity and mortality. Recently, there have been reports indicating the involvement of exosomal circular RNAs (circRNAs) in a range of human disorders and tumor types. However, the role of exosomal circRNAs in bacterial infection remains elusive. Methods We extracted and identified exosomes from the culture medium of PIEC cells infected with or without Glaesserella parasuis . RNA sequencing analysis was performed on the exosomes to screen and identify circRNAs (circHIF1α) associated with Glaesserella parasuis infection. PIEC cells were infected with Staphylococcus aureus or Streptococcus suis 2 to further determine whether exosome-derived circHIF1α was the crucial circHIF1α associated with bacterial infections. The transmission process of exosomes and their circHIF1α between cells was clarified via exosome tracing and co-culture assay. Moreover, the mechanism of circHIF1α being packaged into exosomes was explored, and the effects of exosomes and their circHIF1α on cell proliferation, DNA damage and cell cycle were analyzed. In addition, the binding mode and site of interacting proteins with circHIF1α were further determined. In vivo and in vitro, the role of exosomes and their circHIF1α in host resistance to bacterial infection was confirmed. Results We first discovered a new circHIF1α that was very stable and detectable, encapsulated into exosomes by hnRNPA2B1, and whose expression in exosomes of bacterially infected PIEC cells significantly decreased. Additionally, exosomal circHIF1α reduced bacterial infection both in vitro and in vivo and suppressed the growth of reception cells. Mechanistically, the circHIF1α interacted with the KH domain of IGF2BP3 in an m6A-modified manner, which mediated DNA damage to arrest the cells at the G1/S phase through the interaction between the regulator of Chromosome Condensation 2 (RCC2) and γ-H2AX protein. Exosomal circHIF1α is a unique therapeutic target for bacterial infection since this work highlights its critical function in fighting bacterial infection.
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Journal of Analysis and Testing
·
2024
·
IF 5.5
Xiao Qi,Liu Shuai,Wang Yali,Li Huihao,Yang Mingli,Fang Yi,Chen Sijing,Huang Shan
Human T-cell lymphophilic virus type 1 (HTLV-1), the known retrovirus causing cancer in humans, is closely associated with adult T-cell leukemia/lymphoma and HTLV-1 associated myelopathy/tropical spastic paraparesis. Due to its ability to evade the host's defense mechanisms, early tracking of HTLV-1 becomes crucial. In this study, we integrate λ-Exonuclease (λ-Exo)-assisted target recycling with a terminal deoxynucleotidyl transferase (TdT)-mediated template-free DNA extension process to develop an electrochemical analysis platform for the specific and sensitive detection of HTLV-1 DNA. During the λ-Exo-assisted target recycling, HTLV-1 DNA is recognized by hairpin DNA (Hp-DNA), forming double-stranded DNA (dsDNA) through DNA hybridization. The dsDNA, featuring blunt 5' terminal phosphorylation, is cleaved by λ-Exo, generating abundant short output sequence (sDNA). HTLV-1 DNA is released, initiating a cyclic hybridization-cleavage process. Subsequently, thiol-labelled capture DNA (CP-DNA) assembled on gold electrode surface captures a substantial amount of the generated sDNA, forming CP-DNA-sDNA nanostructures. When TdT and dNTPs are present on the electrode surface, the 3'-OH terminal of sDNA extends to generate long single-stranded DNA (ssDNA) structure. Methylene blue (MB) is selected as the electrochemical signal molecule. MB not only binds with ssDNA but also interacts specifically with dsDNA, resulting in a significantly enhanced electrochemical signal on modified electrode surface. The detection limit of HTLV-1 DNA is as low as 19 amol/L ( S / N = 3) when the two signal amplification strategies are combined. The analysis platform exhibits excellent analytical performance and holds promise as a novel tool for the early tracing and diagnosis of HTLV-1 DNA.
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BIOSENSORS & BIOELECTRONICS
·
2021
·
IF 10.618
Junping Ma,Wushuang Bai,Jianbin Zheng
A self-cleaning electrochemical biosensor based on two-dimensional Cu-porphyrin (Cu-TCPP) metal-organic framework nanofilms , novel super G-quadruplex (G4), and DNA nanomotors was developed for the cyclic detection of Pb 2+ ions. The Cu-TCPP framework with inherent peroxidase activity can create an ultra-thin nanofilm that functioned as a carrier to support the metastable G4 comprising four individual DNA strands. The introduction of Pb 2+ and the intercalation of hemin can help it to form stable G4-hemin DNAzymes, which exhibits strong catalytic H 2 O 2 reduction activity, and its number will be directly related to the amount of the introduced Pb 2+ . Moreover, a DNA nanomotor system is introduced to achieve cyclic detection, and the addition of the fuel DNA strands enables G4 to perform a “complete-dissociation-complete” process for achieving self-cleaning of the electrode interface and the cycle detection of Pb 2+ . The synergistic effects of Cu-TCPP and G4-hemin DNAzymes, which exhibits efficient and catalytic H 2 O 2 reduction, enhance the performance of the electrochemical sensing system. The linear range of this sensor to Pb 2+ is 5 nM–5 μM, and the detection limit is 1.7 nM. Compared with the best system in reported studies, its linear range is five times wider and its detection limit is lower than the previously lowest one. Taking advantage of the Pb 2+ stabilized G4, the proposed sensor can selectively detect Pb 2+ in the presence of other metal ions . The results presented herein comprise a valuable reference for constructing DNA nanoelectronic devices and establish sensitive and cyclic detection of the target and preparing of self-cleaning electrode interfaces.
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BIOSENSORS & BIOELECTRONICS
·
2013
·
IF 5.437
Wenhua Gao,An Zhang,Yunsheng Chen,Zixuan Chen,Yaowen Chen,Fushen Lu,Zhanguang Chen
Biosensor based on DNA hybridization holds great potential to get higher sensitivity as the optimal DNA hybridization efficiency can be achieved by controlling the distribution and orientation of probe strands on the transducer surface. In this work, an innovative strategy is reported to tap the sensitivity potential of current electrochemiluminescence (ECL) biosensing system by dispersedly anchoring the DNA beacons on the gold nanoparticles (GNPs) array which was electrodeposited on the glassy carbon electrode surface , rather than simply sprawling the coil-like strands onto planar gold surface. The strategy was developed by designing a “signal-on” ECL biosensing switch fabricated on the GNPs nanopatterned electrode surface for enhanced ultra-sensitivity detection of Hg 2+ . A 57-mer hairpin-DNA labeled with ferrocene as ECL quencher and a 13-mer DNA labeled with Ru(bpy) 3 2+ as reporter were hybridized to construct the signal generator in off-state. A 31-mer thymine (T)-rich capture-DNA was introduced to form T–T mismatches with the loop sequence of the hairpin-DNA in the presence of Hg 2+ and induce the stem-loop open, meanwhile the ECL “signal-on” was triggered. The peak sensitivity with the lowest detection limit of 0.1 nM was achieved with the optimal GNPs number density while exorbitant GNPs deposition resulted in sensitivity deterioration for the biosensor. We expect the present strategy could lead the renovation of the existing probe-immobilized ECL genosensor design to get an even higher sensitivity in ultralow level of target detection such as the identification of genetic diseases and disorders in basic research and clinical application.
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SENSORS AND ACTUATORS B-CHEMICAL
·
2021
·
IF 7.46
Junyang Wang,Caiyi Du,Peitong Yu,Qian Zhang,Hongxia Li,Chunyan Sun
A label-free and enzyme-free signal-on biosensor for Hg(II) ions was constructed based on T-Hg(II)-T nanoladders and fluorescent DNA-templated silver nanoclusters/graphene oxide nanocomposites (DNA-AgNCs/GO). The T-rich sequences (P1 and P2) were used to capture Hg(II) and form the T-Hg(II)-T nanoladders. C 6 G 5 C 6 was extended at the 5′ end of P1, used as a template for synthesizing fluorescent DNA-AgNCs. In the presence of mercury ions , the T-Hg(II)-T nanoladders significantly enhanced the emission of AgNCs. GO can attract free P1-C 6 G 5 C 6 and P2 that were not involved in the formation of T-Hg(II)-T nanoladders, thus employed as the energy acceptor to quench the background fluorescence of AgNCs. The fluorescence enhancement induced by mercury ions can be used for quantitative detection of Hg(II) over the range 0.1–30 nM with a detection limit as low as 7.35 pM. This method was used to detect Hg(II) in the spiked samples of Songhua River water and grass carp, with the recoveries of 97.61%− 103.79% and 96.52%− 105.58% respectively, consistent with the results from atomic fluorescence spectrometry. Obviously, in this assay, enzyme-free signal amplification and signal identification are integrated in the ingenious design of nucleic acid sequences, thus achieving the label-free fluorescence detection of mercury ions.
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BIOSENSORS & BIOELECTRONICS
·
2016
·
IF 7.476
Fengping Zhan,Xiaolei Liao,Feng Gao,Weiwei Qiu,Qingxiang Wang
A novel electrochemical DNA biosensor has been facilely constructed by in-situ assembly of electroactive 4′-aminobenzo-18-crown-6-copper(II) complex (AbC-Cu 2+ ) on the free terminal of the hairpin-structured molecule beacon. The 3′-SH modified molecule beacon probe was first immobilized on the gold electrode (AuE) surface through self-assembly chemistry of Au–S bond. Then the crow ester of AbC was covalently coupled with 5′–COOH on the molecule beacon, and served as a platform to attach the Cu 2+ by coordination with ether bond (–O–) of the crown cycle. Thus, an electroactive molecule beacon-based biosensing interface was constructed. In comparison with conventional methods for preparation of electroactive molecule beacon, the approach presented in this work is much simpler, reagent- and labor-saving. Selectivity study shows that the in-situ fabricated electroactive molecule beacon remains excellent recognition ability of pristine molecule beacon probe to well differentiate various DNA fragments. The target DNA can be quantatively determined over the range from 0.10 pM to 0.50 nM. The detection limit of 0.060 pM was estimated based on signal-to-noise ratio of 3. When the biosensor was applied for the detection cauliflower mosaic virus 35s (CaMV 35s) in soybean extraction samples, satisfactory results are achieved. This work opens a new strategy for facilely fabricating electrochemical sensing interface, which also shows great potential in aptasensor and immurosensor fabrication.
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Materials Today Bio
·
2025
·
IF 10.2
Qingteng Lai,Yinghui Zhao,Yaqi Dong,Rongzhen Liu,Peng Zhang,Wen Yang,Xiaoshi Zhang,Yi Zhang
Detection of microRNAs (miRNAs) reveals tumor information at the transcriptional level, which is crucial for early diagnosis and prognosis of tumors. However, the rapid detection of miRNAs in biological samples without relying on large equipment remains a significant challenge. In this work, a double-layer NiCo heterostructure (DL-H) for enhancing the reaction efficiency of DNA labels, and dual-target cycling reaction system were developed to improve detection sensitivity and shorten the detection time within 30 min. The DL-H and catechol provide favorable condition for methylene blue (MB) oxidation and single-electron transfer of ferrocene (Fc). The target cycling reaction allows one target to release multiple Fc/MB labels from the electrode surface, enhancing the signal differentiation triggered by the target. The obtained sensor possesses a linear detection range of 10 fM-900 pM for miRNA-574-5p and miRNA-1254. The detection limit is 302 aM for miRNA-574-5p, while 85 aM for miRNA-1254. And the detection results of clinical samples present a strong correlation with the PCR. More importantly, a sensing automatic preparation-detection system was developed based on microfluidic chip, to resolve the reliance on highly trained technicians for the electrochemical sensing preparation. This development provides a valuable foundation for the advancement of portable detection tool of tumor biomarker.
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TALANTA
·
2016
·
IF 4.035
Bei Hua Kang,Zhong Feng Gao,Na Li,Yan Shi,Nian Bing Li,Hong Qun Luo
Silver ions have been widely applied to many fields and have harmful effects on environments and human health. Herein, a label-free optical sensor for Ag + detection is constructed based on thiazole orange (TO) as a fluorescent probe for the recognition of i-motif DNA structure change at neutral pH. Ag + can fold a C-rich single stranded DNA sequence into i-motif DNA structure at neutral pH and that folding is reversible by chelation with cysteine (Cys). The DNA folding process can be indicated by the fluorescence change of TO, which is non-fluorescent in free molecule state and emits strong fluorescence after the incorporation with i-motif DNA. Thus, a rapid, sensitive, and selective method for the detection of Ag + and Cys is developed with a detection limit of 17 and 280 nM, respectively. It is worth noting that the mechanism underlying the increase of the fluorescence of thiazole orange in the presence of i-motif structure is explained. Moreover, a fluorescent DNA logic gate is successfully designed based on the Ag + /Cys-mediated reversible fluorescence changes. The proposed detection strategy is label-free and economical. In addition, this system shows a great promise for i-motif/TO complex to analyze Ag + in the real samples.
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ACS Applied Materials & Interfaces
·
2025
·
IF 8.2
Huajuan Ye,Jinfa Chen,Xianfeng Lv,Wenxin Wu,Zhenli Qiu,Jingyue He,Dage Fan,Ning Li,Bin Han,Junyang Zhuang
The development of photoelectrochemical (PEC) biosensors with enhanced sensitivity and structural simplicity remains a key challenge in biomolecular detection. In this work, we report an unexpected and previously overlooked phenomenon in which DNA aptamers inherently act as amplifiers of PEC signals at semiconductor interfaces. Traditionally regarded solely as passive recognition elements, DNA aptamers─exemplified by the EpCAM-specific SYL3C─were found to markedly increase photocurrent when assembled on graphitic carbon nitride (g-C3N4)-based PEC electrodes. To further enhance interfacial charge transfer, g-C3N4 was covalently functionalized with 1,3,5-benzenetricarboxaldehyde (BTA), forming a donor–acceptor structured semiconductor (g-C3N4–BTA). Density functional theory (DFT) calculations and Mott–Schottky analysis revealed that the lowest unoccupied molecular orbital (LUMO) levels of DNA bases are positioned above the conduction band (CB) edges of both g-C3N4 and g-C3N4–BTA, enabling thermodynamically favorable injection of photoexcited electrons from DNA molecules into the semiconductor CB. This interfacial electron injection, analogous to dye-sensitized solar cells, accounts for the observed PEC signal amplification. Based on this mechanistic understanding, we developed a SYL3C/AuNPs/chitosan/g-C3N4–BTA-modified electrode for ultrasensitive detection of EpCAM-positive exosomes, achieving a detection limit of 988 particles mL–1. Furthermore, the sensor demonstrated robust performance in monitoring phenotypic changes of exosomes secreted by HepG2 cells in response to chemotherapy drug treatment, highlighting its potential for functional exosome analysis in cancer research. This study not only identifies a previously unrecognized inherent property of DNA aptamers to enhance semiconductor photoactivity, but also establishes a minimalist and broadly applicable design principle for constructing high-performance PEC biosensors. The mechanistic insights presented here open new avenues for the rational design of PEC sensing interfaces and extend the utility of DNA aptamers beyond molecular recognition toward active signal amplification.
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