MICROCHEMICAL JOURNAL
·
2018
·
IF 2.746
Lili Yu,Hui Xu,Hou Chen,Shengxiao Zhang,Tingting Jiang,Liangjiu Bai,Wenxiang Wang,Shanmin Gao,Juan Jin
A label-free DNA Y junction sensor for amplified detection of Hg 2+ has been demonstrated by using thymine-Hg 2+ -thymine (T-Hg 2+ -T) recognition, cascade toehold-mediated DNA strand displacement and dye staining double-stranded DNA (dsDNA). In our work, assistant DNA could bind with the first hairpin DNA with toehold domains by the help of thymine-Hg 2+ -thymine (T–Hg 2+ –T) base pairs recognition. The second hairpin DNA with toehold domains will then hybridize with the complementary sequences in first hairpin DNA, which again initiates a new toehold domains to open the third hairpin DNA, triggering a cascade strand displacement reaction. Exonuclease III (Exo III) was added to digest the three hairpin DNA with recessed 3′ ends in the absence of Hg 2+ to reduce the background signal. As a comparison, Exo III is replaced by GO to reduce the background signal because single-strand DNA (ssDNA), three hairpin DNAs with intercalated dye could be adsorbed onto the surface of GO and lead to quenched fluorescence. The method, designed in a simple “mix, incubate, and detect” format, can detect concentrations of Hg 2+ as low as 0.031 nM, and exhibit good selectivity and practical application.
研究领域:
生物化学
·
环境科学
·
分析化学
✓ 已验证
产品关联
DOI
高影响因子
CHEMOMETRICS AND INTELLIGENT LABORATORY SYSTEMS
·
2018
·
IF 2.701
Cai-Xia Shi,Qing Liu,Zeng-Ping Chen,Ru-Qin Yu
Exonuclease III assisted target recycling amplification strategy can be used to enhance the sensitivity of mass spectrometry for the detection of short DNA sequences. However, the distribution pattern of DNA fragments produced by exonuclease III assisted target recycling amplification is generally different for samples with different concentrations of the target DNA sequence, which hinders the extraction of both qualitative and quantitative information of the target DNA from mass spectral measurements using traditional univariate or multivariate models. In this contribution, an advanced model was derived based on a reasonable assumption for the qualitative and quantitative analysis of the mass spectral measurements of DNA fragments produced by exonuclease III-assisted target recycling amplification. Experimental results demonstrated that the integration of exonuclease III assisted target recycling amplification, mass spectrometry and the advanced model could achieve sensitive and accurate quantitative results for a target short DNA sequence in complex biological medium with a detection limit of 50 pM and a mean recovery rate within the range of 89.5%–106.7%. More interestingly, the proposed model could unambiguously identify single nucleotide polymorphisms based on the distribution patterns of residual DNA fragments. Therefore, with the aid of the proposed model, mass spectrometry based on exonuclease III assisted recycling amplification has great potential for the reliable, sensitive, selective, and relatively low-cost detection and quantification of short DNA sequences in clinical diagnosis and biomedical research.
研究领域:
分子生物学
·
临床诊断
·
生物医学研究
✓ 已验证
产品关联
DOI
高影响因子
ANALYST
·
2020
·Vol. 145
·期 8
·2975-2981
·
IF 3.978
Rufeng Zhang,Jie Zhang,Xiaonan Qu,Shasha Li,Yihan Zhao,Su Liu,Yu Wang,Jiadong Huang,Jinghua Yu
DNA walkers, one of the artificial molecular machines which are constructed via smart synthetic DNA, have attracted rapidly growing attention from researchers in the biosensing field. In this work, we design an Exonuclease III (Exo III)-aided target-aptamer binding recycling (ETBR) activated bipedal DNA machine for highly sensitive electrochemical detection of antibiotics. To the best of our knowledge, this is the first time that a bipedal DNA machine has been applied in electrochemical sensing for antibiotics. On the one hand, the bipedal DNA walker exceeds the conventional single swing arm DNA walker in terms of walking efficiency and stability. On the other hand, the ETBR strategy, along with efficient strand displacement amplification via stepwise movement of a bipedal DNA walker significantly promotes the signal amplification efficiency. Under optimal conditions, this bipedal DNA machine possesses a detection limit of 7.1 fM within a linear detection range from 10 fM to 100 pM. Moreover, this electrochemical biosensor is expected to detect a wide variety of analytes using the corresponding target recognition probes. Thus, our proposed strategy offers a highly efficient, stable and practical platform for small molecule analysis.
研究领域:
生物化学
·
分子生物学
·
分析化学
·
纳米技术
✓ 已验证
DOI
PubMed
高影响因子
BIOSENSORS & BIOELECTRONICS
·
2021
·
IF 10.618
Qian Han,Cun Wang,Pingkun Liu,Gui Zhang,Li Song,Yingzi Fu
The excellent characteristics of porphyrins have inspired widespread interest in electrochemiluminescence (ECL). However, the limited ECL intensity and poor stability of porphyrins in aqueous solution are still severely restricted further biological application . Here, we subtly synthesized a functionalized europium and 5,10,15,20-tetrakis (4-carboxyphenyl) porphyrin (TCPP) coordination polymer (Eu-PCP) by a one-step solvothermal method . In sharp contrast to the pristine TCPP, Eu-PCP showed a higher and more stable ECL red-light emission (673 nm) at low potential (−1.1 V, vs Ag/AgCl), which was 7.7-fold higher ECL intensity and 4.6-fold efficiency. In view of the crucial role of mucin 1 (MUC1) in tumor overexpression , it was selected as the target molecule. Combined with exonuclease III (Exo III)-assisted recycling amplification strategy, a ternary ECL biosensor was constructed for the MUC1 detection based on Eu-PCP as a satisfied ECL emitter, gold nanoparticles capped CeO 2 (CeO 2 @Au) as the coreactant accelerator and peroxydisulfate as coreactant. Meanwhile, gold nanoparticles capped MnO 2 (MnO 2 @Au) was used as the quenching probe to achieve a highly sensitive detection of MUC1. The proposed biosensor exhibited a wide linear range from 1 fg mL −1 to 10 ng mL −1 with a low limit of detection (0.32 fg mL −1 ). By changing the corresponding target recognition DNA , this strategy could be expanded to detect other biomarkers.
研究领域:
生物传感
·
分析化学
·
医学诊断
✓ 已验证
DOI
PubMed
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
✓ 已验证
DOI
PubMed
高影响因子
ANALYTICAL CHEMISTRY
·
2016
·
IF 5.886
Wang Ren,Ying Zhang,Hong Guo Chen,Zhong Feng Gao,Nian Bing Li,Hong Qun Luo
A novel signal-on and label-free resonance Rayleigh scattering (RRS) aptasensor was constructed for detection of Hg2+ based on Hg2+-triggered Exonuclease III-assisted target recycling and growth of G-quadruplex nanowires (G-wires) for signal amplification. The hairpin DNA (H-DNA) was wisely designed with thymine-rich recognition termini and a G-quadruplex sequence in the loop and employed as a signal probe for specially recognizing trace Hg2+ by a stable T–Hg2+–T structure, which automatically triggered Exonuclease III (Exo-III) digestion to recycle Hg2+ and liberate the G-quadruplex sequence. The free G-quadruplex sequences were self-assembled into guanine nanowire (G-wire) superstructure in the presence of Mg2+ and demonstrated by gel electrophoresis. The RRS intensity was dramatically amplified by the resultant G-wires, and the maximum RRS signal at 370 nm was linear with the logarithm of Hg2+ concentration in the range of 50.0 pM to 500.0 nM (R = 0.9957). Selectivity experiments revealed that the as-prepared RRS sensor was specific for Hg2+, even coexisting with high concentrations of other metal ions. This optical aptasensor was successfully applied to identify Hg2+ in laboratory tap water and river water samples. With excellent sensitivity and selectivity, the proposed RRS aptasensor was potentially suitable for not only routine detection of Hg2+ in environmental monitoring but also various target detection just by changing the recognition sequence of the H-DNA probe.
研究领域:
分析化学
·
环境科学
·
生物传感技术
✓ 已验证
DOI
PubMed
高影响因子
MICROCHIMICA ACTA
·
2017
·Vol. 185
·期 1
·1-7
·
IF 4.58
Hong Minqiang,Zeng Bihua,Li Mingyu,Xu Xueqin,Chen Guonan
An ultrasensitive conformation-dependent colorimetric assay has been developed for the detection of mercury(II) ions. It is based on the use of exonuclease III (Exo III)-assisted target recycling and gold nanoparticles (AuNPs). In the absence of Hg(II), the hairpin-shaped DNA probe (H-DNA) binds to AuNPs and stabilizes them in solutions of high ionic strength. In the presence of Hg(II), on the other hand, the sticky termini of the H-DNA form a rigid DNA duplex stem with a blunt 3′-terminus. Thus, Exo III is activated as a biocatalyst for selective and stepwise removal of mononucleotides from the 3′-terminus of the H-DNA. As a result, Hg(II) is released from the T−Hg(II)−T complexes. The guanine-rich sequences released from the H-DNA are then self-assembled with potassium ion to form a stable G-quadruplex conformation. In solutions of high ionic strength, this results in aggregation of AuNPs and a color change from red to blue which can be seen with bare eyes. The method is highly sensitive and selective. It has a linear response in the 10 pM to 100 nM Hg(II) concentration range, and the detection limit is as low as 3.2 pM (at an S/N ratio of 3). The relative standard deviation at a level of 0.5 nM of Hg(II) is 4.9% (for n = 10). The method was applied to the detection of Hg(II) in spiked environment water samples, with recoveries ranging from 92% to 106%.
A conformation-dependent colorimetric system was fabricated for label-free detection of mercury(II) by utilizing exonuclease III(Exo III)-assisted target recycling and gold nanoparticles (AuNPs).
研究领域:
分析化学
·
环境科学
·
纳米技术
✓ 已验证
产品关联
DOI
PubMed
高影响因子
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.
研究领域:
生物化学
·
医学诊断
·
纳米技术
✓ 已验证
产品关联
DOI
PubMed
高影响因子
MICROCHIMICA ACTA
·
2024
·Vol. 191
·期 7
·1-12
·
IF 5.3
Tao Chunxu,Wang Junyan,Zhu Ying,Ding Chao,Shen Zhuoyue,Sun Danni,Cao Shanshan,Jiang Xinrong,Li Yaqi,Liu Chang,Zhang Qi,Li Shijie,Zhang Xinyan,Shi Qiaoqiao,Kong Dezhao
A fluorescence biosensor for determination of aflatoxin B 1 (AFB 1 ) based on polydiacetylene (PDA) liposomes and exonuclease III (EXO III)-assisted recycling amplification was developed. The AFB 1 aptamer partially hybridizes with complementary DNA (cDNA), which is released upon recognition of AFB 1 by the aptamer. Subsequently, the cDNA hybridizes with hairpin H to form double-stranded DNA that undergoes digestion by EXO III, resulting in the cyclic release of cDNA and generation of capture DNA for further reaction. The capture DNA then hybridizes with probe modified on PDA liposomes, leading to aggregation of liposomes and subsequent fluorescence production. This strategy exhibited a limit of detection of 0.18 ng/mL within the linear range 1–100 ng/mL with a determination coefficient > 0.99. The recovery ranged from 92.81 to 106.45%, with relative standard deviations (RSD) between 1.73 and 4.26%, for corn, brown rice, peanut butter, and wheat samples. The stability, accuracy, and specificity of the method demonstrated the applicability for real sample analysis. Graphical abstract
研究领域:
食品科学
·
分析化学
·
分子生物学
✓ 已验证
产品关联
DOI
PubMed
高影响因子
Analytical Methods
·
2014
·Vol. 6
·期 12
·4366-4372
·
IF 1.938
Lina Ma,Lan Ma,Min Su,Zhenxin Wang
Herein, a gold nanoparticle (GNP)-based colorimetric aptasensor has been developed for detecting adenosine with exonuclease III (Exo III) assisted recycling amplification. In this aptasensor, two kinds of single-stranded oligonucleotide functionalized GNP probes (Oligo 1-GNPs and Oligo 2-GNPs) are firstly hybridized with one complementary single-stranded oligonucleotide (linker) with the motif of adenosine aptamer, resulting in the formation of GNP aggregates (cross-linked GNPs). In the presence of adenosine, the linker would form a double-stranded DNA (dsDNA) with a recessed 3′-terminus due to its structural switching property, which would lead to disassembly of the cross-linked GNPs. However, upon the addition of Exo III, the dsDNA is enzymatically digested from the 3′-hydroxyl termini, liberating the adenosine. The released adenosine can then interact with another linker and a new round of enzymatic digestion is started. Therefore, remarkable signal amplification is achieved because the disassembly of cross-linked GNPs is strongly dependent on the free adenosine in the reaction mixture. The experimental results demonstrate that the Exo III-assisted GNP-based colorimetric aptasensor is more sensitive (ca. 10 times) than that without Exo III amplification. The Exo III-assisted GNP-based colorimetric aptasensor detects adenosine downs to 5.6 nM, with a linear relationship from 10 nM to 1 mM. Moreover, the Exo III-assisted GNP-based colorimetric aptasensor has been successfully applied to detect Jinshuibao capsule (JSBC) containing adenosine in both a buffer solution and a diluted human serum, demonstrating that this assay has great potential to be employed for detecting adenosine in real samples.
研究领域:
生物化学
·
分析化学
·
纳米技术
✓ 已验证
产品关联
DOI
高影响因子
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.
研究领域:
分析化学
·
生物化学
·
纳米技术
·
分子生物学
✓ 已验证
产品关联
DOI
高影响因子
ANALYST
·
2020
·Vol. 146
·期 4
·1414-1420
·
IF 3.978
Chuanying Qin,Cong Hu,Aimin Yu,Guosong Lai
Herein, we report a Fe3O4@polydopamine (PDA) nanocomposite and exonuclease III (Exo III)-assisted homogeneous fluorescence biosensing method for ultrasensitive detection of kanamycin (Kana) antibiotic. A hairpin DNA containing the Kana-aptamer sequence (HP) was first designed for the highly specific biorecognition of the target analyte. Because of the aptamer biorecognition-induced structural change of HP and the highly effective catalyzed reaction of Exo III, a large amount of fluorophore labels were released from the designed fluorescence DNA probe. During the homogeneous reaction process, the Exo III-assisted dual recycling significantly amplified the fluorescence signal output. Moreover, the excessive probes were easily adsorbed and separated by the Fe3O4@PDA nanocomposite, which decreased the background signal and increased the signal-to-noise ratio. These strategies result in the excellent analytical performance of the method, including a very low detection limit of 0.023 pg mL−1 and a very wide linear range of six orders of magnitude. In addition, this method has convenient operation, excellent selectivity, repeatability and satisfactory reliability, and does not involve the design and utilization of complicated DNA sequences. Thus, it exhibits a promising prospect for practical applications.
研究领域:
分析化学
·
生物传感器
·
分子诊断学
✓ 已验证
DOI
PubMed
高影响因子
危险品化学品经营许可证（带存储）