RSC Advances
·
2016
·Vol. 6
·期 110
·108662-108667
·
IF 3.289
Jin-Xiu Cao,Yong-Sheng Wang,Jin-Hua Xue,Yan-Qin Huang,Ming-Hui Li,Si-Han Chen,Bin Zhou,Xian Tang,Xiao-Feng Wang,Yu-Feng Zhu
We report on a novel and highly sensitive strategy for UO22+ detection. The designed single-labeled hairpin signal probe (SSP) was employed as a reporter, and a multipurpose RNA-cleaving DNAzyme as a target recognition element, catalytic DNAzyme and the primer of substrate fragment recycling amplification. The presence of UO22+ results in the substrate strand in DNAzyme being cleaved at the RNA site, releasing short substrate fragments. The fragment that is complementary partially with SSP could open the hairpin of SSP to form a double-stranded DNA with recessed 3′-termini, which could be selectively digested by exonuclease III. After the double-stranded DNA is fully consumed, the substrate fragment is released. This one could hybridize with another SSP to initiate the next round of digestion by exonuclease III. The recycling of substrate fragments can be repeated multiple times, resulting in the accumulation of the more 6-carboxyfluorescein in the solution along with an amplified fluorescence signal. By this strategy, we achieved hitherto an unreported UO22+ detection limit of 2.4 pM. Therefore, this novel exonuclease III-amplified strategy has great potential for highly sensitive and selective quantification of UO22+ in biomedical research and environmental science fields.
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
TALANTA
·
2017
·
IF 4.162
Shengfeng Huang,Ning Gan,Tianhua Li,You Zhou,Yuting Cao,Youren Dong
An ultrasensitive electrochemical aptasensor for multiplex antibiotics detection based on endonuclease and exonuclease assisted dual recycling amplification strategy was proposed. Kanamycin and chloramphenicol were selected as candidates. Firstly, aptamers of the antibiotics were immobilized on bar A and then binding with their endonuclease labeled complementary DNA strands to construct enzyme-cleavage probes. Secondly, The nano zirconium-metal organic framework (NMOF) particles with 1,4-benzene-dicarboxylate (BDC) as linker was defined as UiO-66. And its updated version, hierarchically porous UiO-66 (HP-UIO-66) decorated with different electroactive materials as signal tags were synthesized. Then they were immobilized on bar B linked by two duplex DNA strands which can be specifically cleaved by corresponding enzyme-cleavage probes in bar A. Once targets were introduced into system, aptamers can capture them and then release enzyme-cleavage probes. In the presence of exonuclease-I, exonuclease assisted target recycling amplification was triggered and more enzyme-cleavage probes were released into solution. The probes can trigger endonuclease assisted recycles and repeatedly cleave their corresponding duplex DNA strands on bar B then released numerous signal tags into supernatant. Thus two recycling amplification was performed in the system. Finally, MB and Fc in the signal tags were detected by square wave voltammetry after removing bar A/B and the current intensities were correspondent with the concentration of KANA and CAP respectively. Under the optimum condition, the limits of detection for the KANA and CAP were 35 fM and 21 fM respectively with a wide linear range from 1 × 10 −4 to 50 nM. This dual recycling amplification detection system exhibited high sensitivities and specificity. It can be easily extended to detect other targets if changing the corresponding aptamers and has potential application values for screening of multiplex antibiotics residues in food safety.
研究领域:
生物化学
·
分析化学
✓ 已验证
产品关联
DOI
PubMed
高影响因子
Analytical Methods
·
2025
·
IF 2.6
Changqing Du,Anran Li,Jinye Qiu,Jiarui Sun,Xiaokun Wang,Wei Xu,Maofeng Zhang,Dapeng Wang
Quantitative mRNA expression levels are essential for differential and clinical diagnoses. In this work, we present an apurinic/apyrimidinic endonuclease 1 (APE1)-assisted SERS sensing platform for rapid and ultrasensitive detection of TK1 mRNA. This platform utilizes a hairpin probe that contains a TK1 mRNA capture sequence along with an apurinic/apyrimidinic (AP) sites, which is specifically recognized and cleaved by APE1. Upon introduction of the target mRNA, hybridization occurs between the mRNA and the AP probe (HAP) within the hairpin structure, forming a doublestranded complex. APE1 then cleaves the HAP at the specific AP sites within the double-stranded complex, leading to a measurable change in the SERS intensity of the FAM-labeled HAP. Based on this principle, the developed SERS sensing platform demonstrates a highly linear response for TK1 mRNA detection across a wider range from 1 fM to 10 nM, with a detection limit (LOD) of 0.2 fM. Moreover, the SERS sensing platform shows great potential for the precise and quantitative detection of TK1 mRNA in human serum, which could be instrumental for mRNA-related research and the early clinical diagnosis of diseases.
✓ 已验证
产品关联
DOI
PubMed
高影响因子
SENSORS AND ACTUATORS B-CHEMICAL
·
2023
·
IF 8.4
Zhimin Li,Keming Wang,Baoting Dou,Zhenbei Li,Yujuan Qi,Po Wang
A growing number of studies have shown the crucial role of microRNA (miRNA) sensing in cancer clinical diagnosis and prognosis research. In this study, a label-free and enzyme-free electrochemical sensor was presented to achieve sensitive determination of miRNA 122, which was constructed based on in situ synthesis of silver nanoclusters (AgNCs) mediated by cascaded recycling amplification. The efficient and cascaded recycling amplification was initiated by the target miRNA 122, resulting in the release of signal probe with rich cytosine bases, which would be loaded on the sensing interface for in situ synthesis of silver nanoclusters , leading to dramatically increased current for quantitative analysis. This method achieved selective and robust miRNA 122 sensing in human serum samples with a detection limit down to 27 aM. The proposed work would provide an innovative method for determining different biomarkers, holding great potential for boosting the development of clinical diagnosis and drug investigation.
研究领域:
生物化学
·
医学诊断
·
纳米技术
✓ 已验证
产品关联
DOI
高影响因子
Materials
·
2018
·Vol. 11
·期 12
·2408
·
IF 2.467
Jingjing Zhang,Jialun Han,Shehong Feng,Chaoqun Niu,Chen Liu,Jie Du,Yong Chen
In this study, a target recycled amplification, background signal suppression, label-free fluorescent, enzyme-free deoxyribonucleic acid (DNA) machine was developed for the detection of adenosine triphosphate (ATP) in human urine. ATP and DNA fuel strands (FS) were found to trigger the operation of the DNA machine and lead to the cyclic multiplexing of ATP and the release of single stranded (SS) DNA. Double-stranded DNA (dsDNA) was formed on graphene oxide (GO) from the combination of SS DNA and complementary strands (CS′). These double strands then detached from the surface of the GO and in the process interacted with PicoGreen dye resulting in amplifying fluorescence intensity. The results revealed that the detection range of the DNA machine is from 100 to 600 nM (R2= 0.99108) with a limit of detection (LOD) of 127.9 pM. A DNA machine circuit and AND-NOT-AND-OR logic gates were successfully constructed, and the strategy was used to detect ATP in human urine. With the advantage of target recycling amplification and GO suppressing background signal without fluorescent label and enzyme, this developed strategy has great potential for sensitive detection of different proteins and small molecules.
研究领域:
生物化学
·
分子生物学
·
分析化学
✓ 已验证
DOI
高影响因子
ANALYST
·
2024
·
IF 3.6
Fengyao Yu,Longyue Qin,Hua Zhang,Longshua Qin,Hao Fan
In this study, we innovatively synthesized bipyridine ruthenium cluster nanosheets (RuMOFNCs), a novel metal–organic framework material that exhibits both fluorescence and electrochemiluminescence. Gold nanoparticles (AuNPs) were anchored onto RuMOFNCs via bipyridine chelation, enhancing optical signals and creating sites for attaching biologically functional probes. We employed tetraferrocene-modified DNA probes, linked via gold–sulfur (Au–S) bonds, to construct a dual-mode fluorescence–electrochemiluminescence biosensor. This sensor, exploiting exonuclease III (Exo III)-mediated cyclic amplification, inhibits the electron transfer from RuMOFNC to tetraferrocene, resulting in amplified fluorescence and electrochemiluminescence signals. The sensor demonstrates exceptional sensitivity for detecting the BRAF gene, with fluorescence and electrochemiluminescence detection limits of 10.3 aM (range: 0.1 fM to 1 nM) and 3.1 aM (range: 1 aM to 10 pM), respectively. These capabilities are attributed to RuMOFNCs’ luminescence properties, tetraferrocene's quenching effect, and the specificity of base pairing. This study's findings hold substantial promise for biomedical research and clinical diagnostics, particularly in precision medicine and early disease detection.
研究领域:
生物医学工程
·
分子诊断
·
纳米技术
✓ 已验证
产品关联
DOI
PubMed
高影响因子
SPECTROCHIMICA ACTA PART A-MOLECULAR AND BIOMOLECULAR SPECTROSCOPY
·
2023
·
IF 4.4
Shunmei Li,Jie Xiang,Fang Yang,Ruo Yuan,Yun Xiang
Methotrexate (MTX) is one of the commonly used therapeutic drugs for treating various tumors and autoimmune diseases. However, high dose usage of MTX may cause severe side effects and the monitoring of MTX is therefore critical. By coupling a new MTX aptamer-based proximity hybridization with polymerization/nicking reaction (PNR) recycling amplifications, we develop here a sensitive and label-free colorimetric approach for MTX detection in diluted human serums. The MTX molecules can bind and switch the conformation of aptamers in the DNA duplex probes to initiate subsequent proximity hybridization-induced PNR recycling processes for the yield of a great deal of G-quadruplexes with the assistance of two single-stranded assistant DNA sequences. Hemin subsequently combines with these G-quadruplexes to produce lots of G-quadruplex/hemin horseradish peroxidase (HRP) mimicking DNAzymes, which then catalyze intensified color transition of the substrate solution to exhibit highly magnified UV–Vis absorption for label-free and ultrasensitive detection of MTX at concentration as low as 5.66 nM in the range of 10 nM to 1 μM. High selectivity of the developed method also enables it to monitor low levels of MTX in diluted serum samples, which offers such a method enormous potentials for convenient and highly sensitive detection of other small molecule drugs for various clinical applications.
研究领域:
生物化学
·
药物分析
·
临床诊断
✓ 已验证
产品关联
DOI
PubMed
高影响因子
BIOCHEMICAL ENGINEERING JOURNAL
·
2022
·
IF 4.446
Ping Ouyang,Yang Qing,Shuhao Zou,Chenxin Fang,Jialun Han,Yuxing Yang,Haiyu Li,Zhencui Wang,Jie Du
It is of great significance to develop a sensitive detection platform for miRNA , a non-invasive biological indicator for the diagnosis, treatment and prognosis monitoring of tumors. In this paper, we design an electrochemical DNA sensor to detect miR-122, based on toehold promoted strand displacement reaction, combined with enzyme assisted cycle amplification. In this strategy, when miR-122 and Exonuclease III (Exo III) coexist in the detection system, the toehold promotes the chain replacement reaction, and the miR-122 hybridizes with a stem-loop DNA, HC-DNA. Simultaneously, Exo III hydrolyzes part of HC-DNA and releases miR-122 and sign-DNA. Then, after multiple enzymatic cyclic amplification reactions (N Cycle), a large amount of Sign-DNA was released. Methylene blue (MB), as the signal molecule, is attached to sign-DNA, close to the surface of the electrode, and resulting in a increased electrochemical signal. Using this method, we can qualitatively analyze the concentration of miR-122 by the amplified electrochemical signals. The detection limit of miR-122 is 0.304 pM, lower than that of the many common large-scale instrumental analysis. Further, the method has good selectivity and anti-interference performance in complex environments. This electrochemical DNA biosensor has potential applications in highly sensitive detection of target molecules even concluding viruses.
研究领域:
生物化学
·
分子生物学
·
分析化学
✓ 已验证
DOI
高影响因子
ANALYST
·
2021
·Vol. 146
·期 8
·2664-2669
·
IF 4.616
Liling Lu,Xiao Han,Jingwen Lin,Yingxin Zhang,Minghao Qiu,Ying Chen,Meijin Li,Dianping Tang
Herein a rapid and sensitive fluorometric bioanalysis platform for mercury(II) (Hg2+) detection was innovatively developed using ultrathin two-dimensional MXenes (Ti3C2) as fluorescence quencher and Hg2+-induced exonuclease III (Exo III)-assisted target recycling strategy for efficient signal amplification. Initially, fluorophore-labeled single-stranded DNA (FAM-labeled probe) can be easily adsorbed onto the surface of ultrathin Ti3C2 nanosheets by hydrogen bonding and metal chelating interaction, and the fluorescence signal emitted by the FAM-labeled probe is quenched strongly owing to the fluorescence resonance energy transfer between the FAM and ultrathin Ti3C2 nanosheets. Upon sensing the target Hg2+, the protruding DNA fragment at the 3′ end of hairpin will hybridize with primer (hairpin-Hg2+-primer), and then further digested by Exo III to produce a probe (nicker). The released target Hg2+ and primer continue to participate in the next recycling, resulting in more hairpin probes becoming nickers. The combination of a large number of nickers and FAM-probe resulted in a significant increase in the fluorescence signal of the system, which was attributed to the fact that the double helix DNA was more rigid and separated from the surface of the ultrathin Ti3C2 nanosheets. The obvious fluorescence signal change of the Ti3C2-based Exo III-assisted target recycling can be accurately monitored by fluorescence spectrometry, which is also proportional to the concentration of Hg2+. Under optimum operating conditions, the peak intensity (520 nm wavelength) of fluorescence increased with increasing Hg2+ within a wide dynamic working range from 0.05 nM to 50 nM (R2 = 0.9913) with a limit of detection down to 42.5 pM. The proposed strategy uses ultrathin MXenes as a platform for binding nucleic acids, which contributes to its potential in nucleic acid hybridization-based biosensing and/or nucleic acid signal amplification bio-applications.
研究领域:
分析化学
·
环境科学
·
纳米技术
·
生物传感
✓ 已验证
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
·
分析化学
·
食品科学
✓ 已验证
DOI
PubMed
高影响因子
TALANTA
·
2024
·
IF 5.6
Lei-Xin Jin,Qi-Meng Wang,Shu-Jiao Zhu,Cai-Lian Zhu,Jian-Jun Sun,Shao-Hua Wu
The flap endonuclease 1 (FEN1) plays a key role in DNA replication and repair, its aberrant expression is associated with tumor development, so it has been recognized as a promising biomarker for a variety of cancers. Here, a novel “turn on” mode gold nanocube-enhanced surface-enhanced Raman scattering (SERS) biosensor was constructed by combining a heated Au electrode (HAuE), exonuclease III (Exo III)-assisted cycle amplification, and gold nanocube (AuNC)-based SERS enhancement to achieve highly sensitive detection of FEN1 activity. The SERS tag was prepared using the Raman reporter modified on the AuNC surface, and the high electromagnetic field provided by the sharp geometric feature of AuNC greatly enhanced the SERS signal. At the same time, HAuE was used to increase the electrode surface temperature and enhance the FEN1 activity, leading to more trigger DNA being cleaved, which was used to initiate the Exo III-assisted cycle amplification. Taking all these advantages, the proposed method possessed high sensitivity and good selectivity, with a low limit of detection (LOD) of 3.19 × 10 −7 U μL −1 . In addition, this method was successfully applied to detect FEN1 activity in real cellular extracts.
研究领域:
生物传感技术
·
分子诊断
·
癌症研究
✓ 已验证
产品关联
DOI
PubMed
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
危险品化学品经营许可证（带存储）