Analytical Methods
·
2016
·Vol. 8
·期 13
·2771-2777
·
IF 1.915
Xue Li,Zhipeng Xie,Wei Wang,Yunlei Zhou,Huanshun Yin,Zhiqing Yang,Shiyun Ai
The DNA isothermal amplification cycle is an effective expansion strategy, which can overcome the inherent limitation of the target to signal ratio of 1 : 1. In this work, we fabricated an electrochemical biosensor for the rapid detection of Dam methyltransferase activity based on the exonuclease III (Exo III) assisted isothermal amplification cycle. First, double stranded DNA (ds-DNA) containing the symmetric sequence of 5′-GATC-3′ was immobilized on the magnetic bead surface. After the ds DNA was methylated by Dam MTase, it could be further cleaved by DpnI and formed a blunt end, which could lead to the digestion of a DNA strand by Exo III and release of a target single stranded DNA (ts-DNA). Then, the ts-DNA can further hybridize with DNA probes (DNA3) immobilized on the AuNPs/Au electrode surface and form a blunt end DNA–DNA duplex. Subsequently, the DNA3 in the DNA–DNA duplex can be digested by Exo III and release ts-DNA. The released ts-DNA can further hybridize with the unhybridized single stranded DNA (ss-DNA) and an isothermal amplification cycle is achieved. The method showed high sensitivity with a low detection limit of 0.031 unit per mL. Moreover, inhibition investigation indicates that Dam MTase activity can be inhibited by using 5-fluorouracil with an IC50 value of 131.22 μM.
研究领域:
生物化学
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分子生物学
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分析化学
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癌症研究
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DOI
高影响因子
CHEMICAL RESEARCH IN CHINESE UNIVERSITIES
·
2022
·Vol. 38
·期 6
·1497-1503
·
IF 2.726
Qin Yang,Yang Yunhan,He Ran,Zhou Laicheng,Zhang Ling
Enzymes containing 3′ → 5′ exonuclease activities play an important role in various key cellular and physiological processes. The development of fluorescence biosensor is an efficient method to detecting enzyme activity. Herein, a fluorescence resonance energy transfer(FRET) “on” and “off” strategy for detecting exonuclease III(Exo III) activity has been developed. We report here that the double-stranded DNA(dsDNA) enables to bind tightly to self-assembled nanosheets of cationic perylene monoimide derivative(PMI-O7) through electrostatic interaction, and the 6-carboxyfluorescein(FAM)-modified dsDNA could be efficiently quenched via FRET between FAM and PMI-O7. Upon the addition of Exo III, the dsDNA will be digested and the FAM fluorophore will be released, resulting in the fluorescence recovery of FAM. This method provides a simple and sensitive biosensor platform with a low detection limit of 0.077 U/mL for Exo III. Importantly, this method exhibits similar and calibration curves for the detection of Exo III in both buffer and fetal bovine serum samples, indicating that this platform has potential to detect Exo III activity in complex samples.
研究领域:
生物化学
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分子生物学
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分析化学
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高影响因子
NEW JOURNAL OF CHEMISTRY
·
2023
·Vol. 47
·期 28
·13228-13234
·
IF 3.3
Yuanyi Wu,Shihan Liu,Yumei Xiao,Qian Li,Qin Xiang,Xianqin Song,Shanglin Wu,Kaiyong Wang,Yi Yang
Accurate detection of DNA is critical for biomedicine and food safety. Layered double hydroxides (LDHs) are a class of two-dimensional layered materials that have received extensive attention in the fields of sensors, catalysis, and materials science. The application of LDHs in DNA sensing in turn represents a fascinating topic. In this study, we found that both 6-carboxyfluorescein (FAM)-modified single-stranded DNA (ssDNA) probes and probe/target double-stranded DNA (dsDNA) duplexes could be adsorbed by LDHs, which resulted in high fluorescence quenching (>95%) for the first time. Based on the extraordinarily high quenching efficiency of LDHs, we developed a fluorescent sensing platform for simple and highly efficient detection of DNA, using Ni/Al-LDHs as fluorescence quenchers. For the selective degradation of dsDNA, exonuclease III was integrated into the sensor. In the sensing platform, the fluorescence of the probes was rapidly quenched by Ni/Al-LDHs. In the presence of the target DNA, dsDNA was formed, which was subsequently selectively degraded by exonuclease III into short fragments, resulting in the maintenance of the fluorescence of FAM labels. The linear range was 10–400 nM. The detection limit was 1.38 nM. This sensor accomplished the detection of target DNA within 50 min. It provides a convenient, rapid and versatile sensing platform for DNA biomarkers.
研究领域:
生物医学工程
·
材料科学
·
分析化学
·
生物传感器
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DOI
高影响因子
ANALYTICAL SCIENCES
·
2024
·
IF 1.8
Liu Xing,Liu Zhen,Pan Yiliang,Xiao Xilin
Construct a sensitive and rapid detection fluorescence biosensor with the assistance of exonuclease III to achieve efficient detection of Hg 2+ . Nucleic acid aptamer H0 specifically recognizes Hg 2+ , and under the action of exonuclease III, H0 bound to Hg 2+ is hydrolyzed, which in turn fails to trigger the catalytic hairpin self-assembly amplification reaction, resulting in a decrease in the number of double-stranded DNA bound to the fluorescent dye SYBR Green I in the solution, and a decrease in the fluorescence intensity. The results showed that the concentration of Hg 2+ showed a good linear relationship with the fluorescence intensity of the sensing system within the range of 3.8–10 pmol/L, and the detection limit was 0.53 pmol/L. The recoveries of Xiangjiang River water used for the analysis of the actual samples were in the range of 99.57–103.58%, and the relative standard deviations of the determined values were 2.4–3.7%. The method is simple, sensitive, specific, cost-effective and can be applied to water samples. Graphical abstract
研究领域:
分析化学
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环境科学
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生物技术
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DOI
PubMed
高影响因子
ACS Applied Materials & Interfaces
·
2016
·
IF 7.145
Xuehong Min,Mengshi Zhang,Fujian Huang,Xiaoding Lou,Fan Xia
Enzyme-assisted detection strategies of microRNAs (miRNAs) in vitro have accomplished both great sensitivity and specificity. However, low expression of miRNAs and a complex environment in cells induces big challenges for monitoring and tracking miRNAs in vivo. The work reports the attempt to carry miRNA imaging into live cells, by enzyme-aided recycling amplification. We utilize facile probes based yellow aggregation-induced emission luminogens (AIEgens) with super photostable property but without quencher, which are applied to monitor miRNAs not only from urine sample extracts (in vitro) but also in live cells (in vivo). The assay could distinguish the cancer patients’ urine samples from the healthy urine due to the good specificity. Moreover, the probe showed much higher fluorescence intensity in breast cancer cells (MCF-7) (miR-21 in high expression) than that in cervical cancer cells (HeLa) and human lung fibroblast cells (HLF) (miR-21 in low expression) in more than 60 min, which showed the good performance and super photostability for the probe in vivo. As controls, another two probes with FAM/Cy3 and corresponding quenchers, respectively, could perform miRNAs detections in vitro and parts of in vivo tests but were not suitable for the long-term cell tracking due to the photobleach phenomena, which also demonstrates that the probe with AIEgens is a potential candidate for the accurate identification of cancer biomarkers.
研究领域:
分子生物学
·
癌症研究
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分析化学
·
生物医学工程
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DOI
PubMed
高影响因子
ANALYTICAL CHEMISTRY
·
2016
·
IF 5.886
Caili Fu,Chang Liu,Ying Li,Yajing Guo,Fang Luo,Peilong Wang,Longhua Guo,Bin Qiu,Zhenyu Lin
Abasic site (AP site) in the triplex structure can recognize specific target with high selectivity. In this study, this character was first applied to develop a simple, sensitive, and selective homogeneous electrochemical biosensor for melamine determination. The assay combines the advantage of the high selectivity of the DNA triplex structure containing an AP site to melamine and high efficiency of exonuclease (Exo) III-assisted recycling amplification. DNA-1 (T1), DNA-2 (T2), poly[dA] probe containing an AP site (8A) and methylene blue-labeled DNA probe (dMB probe) were carefully designed. Melamine can specifically locate in the AP site through hydrogen bonding interaction between thymine and melamine to make T1, T2, and 8A close to each other, therefore, forming a stable T-melamine-T DNA triplex structure. Under the optimal conditions, the differential pulse voltammetric (DPV) response had a linear relationship with the logarithm of melamine concentration in the range of 1 nM∼0.5 μM. The developed biosensor has been successfully applied to detect the migration of melamine from melamine bowl. Result showed that the migration in 4% acetic acid solvent was the largest, which is similar to that detected by high performance liquid chromatography. This homogeneous electrochemical sensor may have a potential prospect in detecting melamine in dairy products and migration of melamine from multicategory food packaging or application materials.
研究领域:
生物化学
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分析化学
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食品科学
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DOI
PubMed
高影响因子
Analytical Methods
·
2025
·
IF 2.6
Lili Zhou,Guangyue Hou,Dan Xi,Yue Guo,Yanping Gou,Hanlin Li,Yaning Zhang,Yongxin Mo,Xiaoli Dai,Rongjin Xu,Hany S. El-Mesery,Wenjie Lu
Mercury ion (Hg²⁺) pollution in tea poses significant risks to human health due to its high toxicity, bioaccumulation, and potential to cause neurological and kidney damage. Sensitive monitoring of Hg²⁺ in tea is therefore critical for food safety and public health. Here, we developed an electrochemical biosensor for ultrasensitive Hg²⁺ assay by integrating thymine-Hg²⁺-thymine (T-Hg²⁺-T) mismatch recognition with exonuclease III (EXO III)-assisted dual signal amplification. The sensing mechanism relies on Hg²⁺-triggered formation of T-Hg²⁺-T structures, which activate EXO III to release Hg²⁺ for cyclic reuse while generating reporter DNA (RDNA). Signal amplification is further enhanced by MOF(Zr)/Th/AuPt nanocomposites, where Zr-based metal-organic frameworks (MOF(Zr)) and AuPt nanoparticles synergistically catalyze thionine (Th) oxidation, producing a strong electrochemical response. This dual-amplification strategy achieves a detection limit of 4.45 pM, surpassing conventional methods. The biosensor demonstrates high specificity against interfering metal ions (e.g., Cd²⁺, Cu²⁺) and reliable performance in real tea samples (93.7%~103.4% recovery), offering a promising tool for monitoring Hg²⁺ contamination in food products.
研究领域:
分析化学
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环境科学
·
食品科学
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产品关联
DOI
PubMed
高影响因子
BIOSENSORS & BIOELECTRONICS
·
2014
·
IF 6.451
Huanshun Yin,Zhiqing Yang,Bingchen Li,Yunlei Zhou,Shiyun Ai
Herein, an electrochemical biosensor was fabricated for DNA demethylase detection based on DNA demethylation triggered endonuclease BstUI and Exonuclease III digestion. After the double-strand DNA was demethylated, it can be further digested by BstUI and formed a blunt end at the electrode surface . Then, the remained fragment of DNA–DNA duplex was further cleaved by exonuclease III and led to increased electrochemical signal. Based on this detection strategy, the biosensor showed high sensitivity with low detection limit of 0.15 ng/mL. Moreover, the developed method also presented high selectivity and acceptable reproducibility. This work provides a novel detection platform for DNA demethylase detection.
研究领域:
生物化学
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分析化学
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分子生物学
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DOI
PubMed
高影响因子
ANALYST
·
2018
·Vol. 143
·期 23
·5771-5778
·
IF 3.864
Xiaolei Song,Yu Wang,Su Liu,Xue Zhang,Haiwang Wang,Jingfeng Wang,Jiadong Huang
In the present work, a simple, rapid, isothermal, and ultrasensitive homogeneous electrochemical biosensing platform for target Hg2+ detection was developed on the basis of an exonuclease III (Exo III)-aided target recycling amplification strategy. In the assay, a label-free hairpin probe (HP1) was ingeniously designed, containing a protruding DNA fragment at the 3′-termini as the recognition unit for target Hg2+. Also, the DNA fragment in the loop region and 5′-termini (Helper) could be used when a secondary target analog is introduced, but it is caged in the stem region of HP1 when without such a target. The produced secondary target Helper opened the methylene blue (MB)-labeled hairpin probe (HP2) and triggered the Exo III cleavage process, accompanied with the secondary target recycling. This accordingly resulted in the autonomous reduction of the electroactive material MB on the electrode, inducing a distinct decrease in the electrochemical signal. The current developed homogeneous strategy provides a means for the ultrasensitive electrochemical detection of Hg2+ down to the 227 pM level, with high selectivity. It could be further used as a general autocatalytic and homogeneous strategy toward the detection of a wide spectrum of analytes and may be associated with more analytical techniques.
研究领域:
分析化学
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环境科学
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生物化学
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DOI
PubMed
高影响因子
SCIENCE OF THE TOTAL ENVIRONMENT
·
2023
·
IF 9.8
Kai Wang,Han Yan,Baoshan He,Lingling Xie,Renli Liu,Min Wei,Huali Jin,Wenjie Ren,Zhiguang Suo,Yiwei Xu
Vomitoxin (DON) residues in grains are of great concern to public health. Herein, a label-free aptasensor was constructed to detect DON distributed in grains. Cerium-based metal-organic framework composite gold nanoparticles ( [email protected] ) were used as substrate materials to facilitate electron transfer and provided more binding sites for DNA. The separation of DON-aptamer (Apt) complex and cDNA was achieved by magnetic separation technique based on magnetic beads (MBs), ensuring the specificity of the aptasensor. Exonuclease III (Exo III)-assisted cDNA cycling process strategy would be triggered when cDNA was separated and introduced to the sensing interface for further signal amplification. Under optimal conditions, the constructed aptasensor presented a wide detection range from 1 × 10 −8 mg·mL −1 to 5 × 10 −4 mg·mL −1 for DON, and the detection limit was 1.79 × 10 −9 mg·mL −1 , including a satisfactory recovery in cornmeal sample spiked with DON. The results showed that the proposed aptasensor had high reliability and promising application potential in detecting DON.
研究领域:
分析化学
·
食品科学
·
生物传感器
·
毒理学
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产品关联
DOI
PubMed
高影响因子
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
·
肿瘤学
·
转化医学
·
生物传感
·
临床检验
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DOI
PubMed
高影响因子
BIOSENSORS & BIOELECTRONICS
·
2014
·
IF 6.451
Wang Ren,Zhong Feng Gao,Nian Bing Li,Hong Qun Luo
This work reported a novel, ultrasensitive, and selective platform for electrochemical detection of DNA , employing an integration of exonuclease III (Exo-III) assisted target recycling and hybridization chain reaction (HCR) for the dual signal amplification strategy. The hairpin capture probe DNA (C-DNA) with an Exo-III 3′ overhang end was self-assembled on a gold electrode. In the presence of target DNA (T-DNA), C-DNA hybridized with the T‐DNA to form a duplex region, exposing its 5′ complementary sequence (initiator). Exo-III was applied to selectively digest duplex region from its 3-hydroxyl termini until the duplex was fully consumed, leaving the remnant initiator. The intact T-DNA spontaneously dissociated from the structure and then initiated the next hybridization process as a result of catalysis of the Exo-III. HCR event was triggered by the initiator and two hairpin helper signal probes labeled with methylene blue , facilitating the polymerization of oligonucleotides into a long nicked dsDNA molecule. The numerous exposed remnant initiators can trigger more HCR events. Because of integration of dual signal amplification and the specific HCR process reaction, the resultant sensor showed a high sensitivity for the detection of the target DNA in a linear range from 1.0 fM to 1.0 nM, and a detection limit as low as 0.2 fM. The proposed dual signal amplification strategy provides a powerful tool for detecting different sequences of target DNA by changing the sequence of capture probe and signal probes, holding a great potential for early diagnosis in gene-related diseases.
研究领域:
生物化学
·
分析化学
·
纳米技术
·
分子生物学
·
生物医学工程
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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）