SENSORS AND ACTUATORS B-CHEMICAL
·
2018
·
IF 5.667
Chen Li,Xuejuan Chen,Zhe Zhang,Jilin Tang,Bailin Zhang
Aptamer-based microcantilever (ML) array sensor is a recently development method for the analysis of various molecules with free-label, high sensitivity and selectivity. Since its signal-transducing principle is the changes of surface stress or resonance frequency caused by molecules binding on the surface, there have difficulties in determining some low molecular weight substances. Herein, a novel detecting dopamine (DA) method based on aptamers functionalized-ML array using gold nanoparticle (NP)-DNA conjugates as the signal-amplifying strategy was developed. In this method, thiolated DA aptamers were self-assembled on the surface of ML array and then Au NP-DNA conjugates which contained a stretch of complementary sequences for DA aptamers were hybridized to DA aptamers. When DA sample interacted with its aptamer and replaced the Au NP-DNA conjugates, the surface stress of ML array was enhanced about fifteen times and resulted in much more deflection. The deflection was linearly proportional to DA concentration in the range from 0.5 to 4 μM with a lowest limit of detection 77.3 nM ( R 2 = 0.993). Some structural and functional DA analogues did not cause interference for the detection of DA. These results proved that the functionalized-ML array sensor based on Au NP-DNA conjugates could be used as a promising tool to detect low-weight molecules with high sensitivity and selectivity.
研究领域:
生物传感技术
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分析化学
·
纳米技术
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DOI
高影响因子
MICROCHEMICAL JOURNAL
·
2022
·
IF 5.304
Wanling Cui,Xiaoyang Fan,Congcong Shen,Qingshuai Chen,Xiumei Zhang,Enguang Lv,Chonghui Li,Qiang Li,Rui Wang,Shicai Xu,Jihua Wang
T4 polynucleotide kinase (T4 PNK) and T4 DNA ligase are two important repair enzymes that play a significant role in maintaining the stability and integrity of genome. Here, we proposed a multiple primers-mediated exponential rolling circle amplification strategy for label-free and highly sensitive fluorescent analysis of T4 PNK and T4 DNA ligase activity. Firstly, we designed a target strand that contained two C-rich sequences, a complementary sequence of the primer, and three cleavage sites of Nt.BbvCI scattered among the three sequences. The 5′-hydroxyl terminus of target strand was catalyzed by T4 PNK to form a phosphorylated target strand. After the phosphorylated target strand was combined with the primer, T4 DNA ligase catalyzed the formation of circular DNA. Then, the primer triggered an exponential rolling circle amplification reaction along the circular DNA template, exponentially generating G-rich sequences. N-methyl mesoporphyrin IX as signal molecules specifically bound to the G-quadruplexs folded by G-rich sequences, generating a detectable fluorescent signal. Benefiting from exponential signal growth, this strategy showed extremely high detection sensitivity with detection limits of 8.11 × 10 −5 U mL −1 of T4 PNK and 3.07 × 10 −4 U mL −1 of T4 DNA ligase, respectively. In addition, the proposed strategy exhibited excellent inhibitor evaluation effects for two repair enzymes, and could detect T4 PNK in HeLa cell lysate. The obtained results showed that the proposed strategy had excellent application prospects in disease diagnosis and treatment evaluation.
研究领域:
生物化学
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分子生物学
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医学诊断
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DOI
高影响因子
Scientific Reports
·
2016
·Vol. 6
·期 1
·1-8
·
IF 5.228
Lan Jianming,Liu Yingxin,Li Li,Wen Fadi,Wu Fang,Han Zhizhong,Sun Weiming,Li Chunyan,Chen Jinghua
High-sensitivity detection of trace amounts of c-erbB-2 oncogene was reported to be equal to or surpass the ability of CA 15-3 for early diagnosis and/or follow-up recurrent screening of breast cancer. Therefore, in the current study, by using upconversion nanoparticles (UCNPs), rare earth-doped NaYF 4 :Yb 3+ /Er 3+ as the luminescent labels, a upconversion luminescent (UCL) biosensor based on dual-signal amplification of exonuclease III (ExoIII)-assisted target cycles and long-range self-assembly DNA concatamers was developed for the detection of c-erbB-2 oncogene. The proposed biosensor exhibited ultrasensitive detection with limit as low as 40 aM, which may express the potential of being used in trace analysis of c-erbB-2 oncogene and early diagnosis of breast cancer.
研究领域:
生物传感
·
癌症诊断
·
分子生物学
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DOI
PubMed
高影响因子
Biosensors-Basel
·
2023
·Vol. 13
·期 9
·875
·
IF 5.4
Ziang Wu,Roujuan Kou,Kun Ni,Rui Song,Yuxuan Li,Tianhu Li,Hao Zhang
DNA ligases are essential enzymes involved in DNA replication and repair processes in all organisms. These enzymes seal DNA breaks by catalyzing the formation of phosphodiester bonds between juxtaposed 5′ phosphate and 3′ hydroxyl termini in double-stranded DNA. In addition to their critical roles in maintaining genomic integrity, DNA ligases have been recently identified as diagnostic biomarkers for several types of cancers and recognized as potential drug targets for the treatment of various diseases. Although DNA ligases are significant in basic research and medical applications, developing strategies for efficiently detecting and precisely quantifying these crucial enzymes is still challenging. Here, we report our design and fabrication of a highly sensitive and specific biosensor in which a stable DNA hairpin is utilized to stimulate the generation of fluorescence signals. This probe is verified to be stable under a wide range of experimental conditions and exhibits promising performance in detecting DNA ligases. We anticipate that this hairpin-based biosensor will significantly benefit the development of new targeting strategies and diagnostic tools for certain diseases.Keywords:DNA ligases;extraordinarily stable hairpins;biomarker detection;fluorescence probes;molecular sensing
研究领域:
分子生物学
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医学诊断
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生物化学
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产品关联
DOI
PubMed
高影响因子
ANALYST
·
2013
·Vol. 138
·期 17
·4737-4740
·
IF 3.969
Chenchen Ge,Junhua Chen,Wei Wu,Zhiyuan Fang,Lingbo Chen,Qi Liu,Lin Wang,Xuerong Xing,Lingwen Zeng
An enzyme-free and label-free fluorescence turn on biosensor for amplified copper(II) ion (Cu2+) detection has been constructed based on self-assembled DNA concatamers and Sybr Green I. This assay is simple, inexpensive and sensitive, enabling quantitative detection of as low as 12.8 pM Cu2+.
研究领域:
分析化学
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生物传感器
·
纳米技术
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DOI
PubMed
高影响因子
SENSORS AND ACTUATORS B-CHEMICAL
·
2018
·
IF 5.667
Qian Kuang,Chunli Li,Zhiwei Qiu,Guifen Jie,Shuyan Niu,Tingyu Huang
In this work, a target-triggered three-way junction (3-WJ) structure was combined with enzyme-powered cascade amplification strategy for ultrasensitive electrochemiluminescence (ECL) detection of microRNA via free-running DNA walker. In the presence of target microRNA-21 (miRNA-21), a three-way junction structure was formed to trigger enzyme-aided multiple DNA amplification, and thus exponentially amplified triggers were generated. Then, the trigger DNA could hybridize with the blocking DNA on the electrode to release walker DNA, which specifically combined with the quantum dots (QDs)-DNA probe, and formed a recognition site for Nt.BbvCI. The movement of DNA walker is powered by the nicking endonuclease that cleaves specific QDs-DNA probes on the track. Thus one walker DNA can automatically cleave multiple QDs probes on the electrode during the movement, resulting in significantly amplified changes of ECL signal, which could be used for the highly sensitive detection of target miRNA-21. Under the optimal conditions, the ECL signal change is linear with the concentration of miRNA-21 in the range from 10 −14 M to 10 -7 M with a detection limit of 1.5 fM. This newly established strategy could achieve rapid, isothermal, and homogeneous signal amplification for specific nucleic acids in complicated biomatrix, which hold great potential for application in early clinical diagnosis.
研究领域:
生物化学
·
分子生物学
·
分析化学
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产品关联
DOI
高影响因子
SENSORS AND ACTUATORS B-CHEMICAL
·
2022
·
IF 9.221
Mei Wang,Hanfeng Cui,Nian Hong,Qingxia Shu,Xinru Wang,Yuping Hu,Guobing Wei,Hao Fan,Jing Zhang
In this paper, we first synthesized a novel tetraferrocene signal marker. Then a reagentless triplex DNA junctions-based electrochemical DNA sensor using signal amplification strategy of CHA and tetraferrocene was constructed. Tetraferrocene labeled hairpin probes (H1 and H2) were integrated together to form triplex DNA junctions through connecting chain (C1). It enabled catalytic hairpin assembly (CHA) reaction to be initiated without the addition of exogenous probe during the detection process, and the conformational change of the target caused the change of electrochemical signal. Combined with tetraferrocene and CHA strategies, the sensor achieved double signal amplification with a detection limit as low as 0.085pM. In addition, the sensor was successfully applied to mRNA detection in cells with high sensitivity and reagentless operation, providing possibilities for in vivo testing.
研究领域:
生物化学
·
分子生物学
·
分析化学
·
生物技术
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DOI
高影响因子
SENSORS AND ACTUATORS B-CHEMICAL
·
2023
·
IF 8.4
Qing Han,Xinlan Bian,Yue Chen,Bingzhi Li
Flap endonuclease I (FEN1) is a structure-specific nuclease that participates in DNA repair, and its overexpression is related to an increased risk of cancer. Currently, few FEN1 assays can meet the requirements for diagnostic purposes due to their high complexity, low specificity, and limited sensitivity. Herein, we developed a biosensor adopting synthetic biology strategy, called CLAIM (cleavage-ligation-activation-involved monitoring) for FEN1 quantification. CLAIMintegrates ligation-based T7 promoter repair, RNA generation, and CRISPR/Cas13a. FEN1 cleavages the DNA probe with a 5′-flap to generate a nick site that can be ligated to form a T7 promoter. Subsequent transcription produces abundant RNA that can activate Cas13a, generating a fluorescence signal due to the trans -cleavage of Cas13a to a fluorogenic reporter. CLAIM was able to detect FEN1 with low femtomolar sensitivity, and its accuracy was validated using samples of cancer and non-tumorigenic cell lines. Interestingly, we found that the utilization of thermostable T4 DNA ligase is critical to make the cascade of FEN1 cleavage, ligation, and transcription take place in one-pot. CLAIM was also repurposed for validating FEN1 inhibitors, indicating its potential in drug screening. This work provides a diagnostic tool for FEN1 analysis and opens up opportunities for FEN1 biosensing and related drug discovery.
研究领域:
分子生物学
·
合成生物学
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癌症诊断
·
生物技术
✓ 已验证
DOI
高影响因子
ANALYTICAL CHEMISTRY
·
2018
·
IF 6.042
Xudong Wang,Jun Dai,Xuehong Min,Zhihua Yu,Yong Cheng,Kaixun Huang,Juliang Yang,Xiaoqing Yi,Xiaoding Lou,Fan Xia
Detection of an ultralow concentration of mRNA is important in the prognosis of gene-related diseases. In this study, a DNA-conjugated amphiphilic aggregation-induced emission probe (TPE-R-DNA) was synthesized for cancer tissue imaging and prognosis analysis based on an exonuclease III-aided target recycling technique. TPE-R-DNA comprise two components: a hydrophobic component that serves as the “turn-on” long wavelength fluorescence imaging agent (TPE-R-N3); and a hydrophilic single DNA strand (Alk-DNA) which acts as specific recognition part for target mRNA. In the absence of target mRNA, TPE-R-DNA had almost no fluorescence because of its high water solubility. Conversely, the TPE-R-DNA was digested by exonuclease III (Exo III) in the presence of MnSOD mRNA to release the hydrophobic fluorogens (TPE-R-AT). Subsequently, TPE-R-AT formed aggregates, and therefore, fluorescence signal was distinctly observed. For the first time, the structure of the hydrolysis product (TPE-R-AT), containing two bases A and T, was proved by the mass spectrum (MS) and high-performance liquid chromatography (HPLC). Moreover, the detection limit toward mRNA could be achieved in as low as 0.6 pM. Furthermore, the fluorescent signal can be used to confirm the MnSOD mRNA expression level in cancer tissue. The MnSOD mRNA expression in renal cancer was lower than in renal cancer adjacent tissue. In particular, the expression level was analyzed to predict prognosis of cancer patients. Our results demonstrate that a shorter survival time was evident among patients in lower MnSOD mRNA expression. Thereby, it indicates great potential for the development of an ultrasensitive biosensing platform for the application in disease prognosis.
研究领域:
癌症研究
·
分子生物学
·
生物医学工程
·
分析化学
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产品关联
DOI
PubMed
高影响因子
TALANTA
·
2018
·
IF 4.244
Qiao Liu,Xing-Li Xie,Chang-Jie Mao,Jing-Shuai Chen,He-Lin Niu,Ji-Ming Song
A highly selective DNA-based electrochemiluminescence (ECL) based biosensor is described for the detection of human IgG. It is exploiting the effect of steric hindrance that affects the strength of the ECL signal in the presence of IgG. Digoxin-linked signaling DNA was specifically bound to IgG, and this causes steric hindrance which limits the ability of DNA to hybridize with capturing DNA attached to a gold electrode. Europium (II) doped CdSe quantum dots were covalently linked to the DNA in order to generate the ECL signal. Using this steric hindrance hybridization method, the ECL signal of the biosensor were proportional to the concentration of IgG with a wide linear range and a 14 pM detection limit. Conceivably, the method can be expanded to the detection of a wide range of proteins for which homologous recognition elements are available.
研究领域:
生物化学
·
分析化学
·
生物传感器
·
免疫学
✓ 已验证
产品关联
DOI
PubMed
高影响因子
MICROCHEMICAL JOURNAL
·
2021
·
IF 4.821
Qian Zhao,Guangyin Zhang,Dai Lu,Kejun Feng,Xingbo Shi
Herein, an ultra-sensitive aptasensor based on dual-enzyme mediated cascade- signal amplification (dubbed Demcsa aptasensor) for ampicillin (AMP) detection is proposed. The sensor’s activity involves two major steps, i.e., DNA ligase-assisted Exonuclease III (Exo III) catalyzed degradation reaction in a homogeneous solution, signal generation/amplification through the biotin-streptavidin linkage and hybridization chain reaction (HCR) on a 96-well polystyrene plate. Firstly, a detection probe was formed via the hybridization reaction between biotinylated AMP aptamer (bio-AP), HCR initiator strand (H strand) and complementary DNA (cDNA). As a result of the dual-enzyme mediated degradation, a long APH strand was formed via the DNA ligase-catalyzed ligation of bio-AP and the H strand, which served as integrated initiator to trigger the HCR reaction, thereby producing a long double-stranded DNA polymer. Subsequently, the SYBR Green I, as fluorescent dye for DNA, was added and a strong fluorescence signal was generated. In the presence of AMP, the function of DNA ligase was effectively suppressed due to the higher affinity of target-AP strand complex. As a result, the HCR reaction could not be triggered without generating a fluorescence signal, because the bio-AP cannot ligate to the H strand even when captured on the plate. The cascade- signal amplification of biotin-streptavidin linkage and the HCR reaction endowed the Demcsa aptasensor with a limit of detection (LOD) of aM, which was comparable or superior to those of the previously reported sensors. The Demcsa aptasensor has been used for the screening of AMP in food samples with favorable recoveries. The results of this work pave a new, universal way for developing efficient sensors for the detection of other trace food hazards.
研究领域:
分析化学
·
生物传感器
·
食品科学
·
分子生物学
✓ 已验证
产品关联
DOI
高影响因子
BIOSENSORS & BIOELECTRONICS
·
2014
·
IF 6.451
Lu Lu,Jing Cao Si,Zhong Feng Gao,Yu Zhang,Jing Lei Lei,Hong Qun Luo,Nian Bing Li
A dual strategy that combines the adenosine triphosphate (ATP)-dependent enzymatic ligation reaction with self-cleaving DNAzyme-amplified electrochemical detection is employed to construct the biosensor. In this design, the methylene blue-labeled hairpin-structured DNA was self-assembled onto a gold electrode surface to prepare the modified electrode through the interaction of Au–S bond. In the procedure of ATP-dependent ligation reaction, when the specific cofactor ATP was added, the two split oligonucleotide fragments of 8-17 DNAzyme were linked by T4 DNA ligase and then released to hybridize with the labeled hairpin-structured DNA substrate. The linked 8-17 DNAzyme catalyzes the cleavage of the hairpin-structured substrate by the addition of Zn 2+ , causing the methylene blue which contains high electrochemical activity to leave the surface of the gold electrode, therefore generating a dramatic decrease of electrochemical signal. The decrease of peak current was readily measured by square wave voltammetry and a relatively low detection limit (0.05 nM) was obtained with a linear response range from 0.1 to 1000 nM. By taking advantage of the highly specific cofactor dependence of the DNA ligation reaction, the proposed ligation-induced DNAzyme cascades demonstrate ultrahigh selectivity toward the target cofactor ATP. A catalytic and molecular beacons strategy is further adopted to amplify the electrochemical signal detection achieved by cycling and regenerating the 8-17 DNAzyme to realize enzymatic multiple turnover, thus one DNAzyme can catalyze the cleavage of several hairpin-structured substrates, which improves the sensitivity of the newly designed electrochemical sensing system.
研究领域:
生物化学
·
电化学
·
生物传感器
·
纳米技术
✓ 已验证
DOI
PubMed
高影响因子
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