BIOSENSORS & BIOELECTRONICS
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2025
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IF 10.5
Jing Peng,Simin Liu,Ziyue Qiu,Luyin Zhang,Ziyan Dong,Qianying Zhou,Ting Huang,Yanfei Zhang,Jingyu Shi,Jin-Xiang Chen,Zong Dai,Jun Chen
miRNA is an effective disease diagnostic biomarker. Developing efficient miRNA in living cells analysis methods, holds significant importance for disease diagnosis and understanding disease progression. Strategies combining DNA nanostructures and enzyme-free amplification have been developed for highly sensitive and rapid imaging of miRNA. However, due to nonspecific amplification of enzyme-free amplification inherent in the probe assembly process and during transfection, these strategies suffer from significant background signals and reduced sensitivity, limiting their further application. To overcome these limitations, we have developed a DNA tetrahedral-based and photocontrolled intramolecular CHA strategy (TCHA-P) to achieve spatiotemporal, highly sensitive, and accurate imaging of miRNA in living cells. In this strategy, the DNA tetrahedron not only serves as a transfection vector but also enhances the local concentration of reactant probes, boosting the reaction kinetics and sensitivity of CHA. Additionally, the light-controlled group (PC-linker) can trigger CHA at selected times and locations, enhancing the spatiotemporal precision of miRNA imaging and ensuring high sensitivity and accuracy by eliminating nonspecific amplification during the assembly and transfection processes. Using miR-10b as a proof-of-concept model, the proposed strategy demonstrated higher signal-to-noise ratio compared to traditional CHA, with a detection limit of 28 pM. Living cell imaging experiments showed that the strategy could effectively differentiate the expression of miR-10b between normal breast cells and breast cancer cells, providing high imaging contrast. The strategy holds promise for broadening the path of nucleic acid amplification-based imaging and is expected to become an effective tool in nucleic acid-based disease diagnosis, treatment, and drug development.
研究领域:
分子生物学
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纳米生物学
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医学诊断
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产品关联
DOI
PubMed
高影响因子
ACS Applied Materials & Interfaces
·
2023
·
IF 9.5
Shaoqiong Feng,Hanjun Chen,Ziao Hu,Tingting Wu,Zhihong Liu
MicroRNA (miRNA) is a promising biomarker for the diagnosis, monitoring, and prognostic evaluation of diseases, especially cancer. The existing miRNA detection methods usually need external instruments for quantitative signal output, limiting their practical applications in point-of-care (POC) settings. Here, we propose a distance-based biosensor through a responsive hydrogel, in combination with a CRISPR/Cas12a system and target-triggered strand displacement amplification (SDA) reaction for visual quantitative and sensitive measurement of miRNA. The target miRNA is first converted into plenty of double-stranded DNA (dsDNA) via target-triggered SDA reaction. Then, the dsDNA products trigger the collateral cleavage activity of CRISPR/Cas12a, leading to the release of trypsin from magnetic beads (MBs). The released trypsin can hydrolyze gelatin, and hence the permeability of gelatin-treated filter paper is increased, resulting in a visible distance signal on a cotton thread. Using this system, the concentration of the target miRNA can be quantified visually without any assistance of instruments, and a detection limit of 6.28 pM is obtained. In addition, the target miRNA in human serum samples and cell lysates can also be detected accurately. Owing to the characteristics of simplicity, sensitivity, specificity, and portability, the proposed biosensor provides a new tool for miRNA detection and holds great promise in POC applications.
研究领域:
生物技术
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分子诊断
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医学检验
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DOI
PubMed
高影响因子
TALANTA
·
2018
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IF 4.244
Xiaojuan Liu,Ningning Xu,Panpan Gai,Feng Li
Since melamine is a strong hazard to human health, the development of new methods for highly sensitive detection of melamine is highly desirable. Herein, a novel fluorescent biosensing strategy was designed for sensitive and selective melamine assay based on the recognition ability of abasic (AP) site in triplex towards melamine and signal amplification by Mg 2+ -dependent DNAzyme. In this strategy, the melamine-induced formation of triplex DNA was employed to trigger the strand displacement reaction (SDR). The SDR process converted the specific target recognition into the release and activation of Mg 2+ -dependent DNAzyme, which could catalyze the cleavage of fluorophore/quencher labeled DNA substrate (FQ), resulting in a significantly increased fluorescent signal. Under the optimal conditions, the fluorescent signal has a linear relationship with the logarithm of the melamine concentration in a wide range of 0.005–50 μM. The detection limit was estimated to be 0.9 nM (0.1ppb), which is sufficiently sensitive for practical application. Furthermore, this strategy exhibits high selectivity against other potential interfering substances, and the practical application of this strategy for milk samples reveals that the proposed strategy works well for melamine assay in real samples. Therefore, this strategy presents a new method for the sensitive melamine assay and holds great promise for sensing applications in the environment and the food safety field.
研究领域:
生物化学
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食品安全
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分析化学
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DOI
高影响因子
ANALYTICAL BIOCHEMISTRY
·
2023
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IF 2.9
Zongkang Guo,Xueting Fan,Xiaoli Wang,Zhemin Zhou,Yuting Zhang,Nandi Zhou
Mec A, as a representative gene mediating resistance to β‐lactam antibiotics in methicillin-resistant Staphylococcus aureus (MRSA), allows a new genetic analysis for the detection of MRSA. Here, a sensitive, prompt, and visual colorimetry is reported to detect the Mec A gene based on toehold‐mediated strand displacement (TMSD) and the enrichment effect of graphene oxide (GO). The Mec A triggers to generate the profuse amount of signal units of single-stranded DNA (SG) composed of a long single‐stranded base tail and a base head: the tail can be adsorbed and enriched on the surface of GO; the head can form a G quadruplex structure to exert catalytic function towards 2,2′‐azino‐bis (3‐ethylbenzthiazoline‐6‐sulphonic acid). Therefore, through the enrichment effect of GO, the signal units SG reflects different degrees of signal amplification on different substrates (such as aqueous solution or filter membrane). This strategy demonstrates a broad linear working range from 100 pM to 1.5 nM (solution) and 1 pM to 1 nM (filter membrane), with a low detection limit of 39.53 pM (solution) and 333 fM (filter membrane). Analytical performance in real samples suggests that this developed colorimetry is endowed with immense potential for clinical detection applications.
研究领域:
生物化学
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医学诊断
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纳米材料科学
✓ 已验证
产品关联
DOI
PubMed
高影响因子
BIOELECTROCHEMISTRY
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2022
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IF 5.76
Jinzhi Liu,Mengyan Wang,Chi Zhang,Gaiping Li,Qi Shen,Lina Zou
Herein, a “signal-on” electrochemical aptasensor based on dual-output toehold mediated strand displacement reaction (TMSDR) was developed to detect cytokeratin fragment antigen 21–1 (CYFRA 21–1). First, the methylene blue (MB) labeled signal probe (SP) was combined with a long template strand (TS) to form the three-strands duplex, which was immobilized on magnetic beads. The sequence of TS was innovatively designed with two toehold zones. Thus, with the presence of target DNA and Fuel strand (F), double SP and target were released through TMSDR. Besides, the released target could participate in the next cycle, achieving the further amplification. The released SP was selected by magnetic separation, then added to the gold electrode (GE) to hybridize with the hairpin DNA. Ultimately, the target can be detected by recording the current response of the MB. Under optimized conditions, the response peak current was linear with the concentration of CYFRA 21–1 in range from 10 pM to 1 μM with detection limit of 8.079 pM. The proposed signal amplification strategy was enzyme-free in homogenous solution and the using of magnetic beads improved the recognition efficiency and simplified the experimental steps, which endowed the sensor with super-sensitivity.
研究领域:
生物化学
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医学检测
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分子生物学
✓ 已验证
DOI
PubMed
高影响因子
MICROCHIMICA ACTA
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2018
·Vol. 185
·期 2
·1-7
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IF 5.705
Ge Jia,Bai Dong-Mei,-Geng Xin,Hu Ya-Lei,Cai Qi-Yong,Xing Ke,Zhang Lin,Li Zhao-Hui
The authors describe a fluorometric method for the quantitation of nucleic acids by combining (a) cycled strand displacement amplification, (b) the unique features of the DNA probe SYBR Green, and (c) polydopamine nanotubes. SYBR Green undergoes strong fluorescence enhancement upon intercalation into double-stranded DNA (dsDNA). The polydopamine nanotubes selectively adsorb single-stranded DNA (ssDNA) and molecular beacons. In the absence of target DNA, the molecular beacon, primer and SYBR Green are adsorbed on the surface of polydopamine nanotubes. This results in quenching of the fluorescence of SYBR Green, typically measured at excitation/emission wavelengths of 488/518 nm. Upon addition of analyte (target DNA) and polymerase, the stem of the molecular beacon is opened so that it can bind to the primer. This triggers target strand displacement polymerization, during which dsDNA is synthesized. The hybridized target is then displaced due to the strand displacement activity of the polymerase. The displaced target hybridizes with another molecular beacon. This triggers the next round of polymerization. Consequently, a large amount of dsDNA is formed which is detected by addition of SYBR Green. Thus, sensitive and selective fluorometric detection is realized. The fluorescent sensing strategy shows very good analytical performances towards DNA detection, such as a wide linear range from 0.05 to 25 nM with a low limit of detection of 20 pM.
Schematic of a fluorometric strategy for highly sensitive and selective determination of nucleic acids by combining strand displacement amplification and the unique features of SYBR Green I (SG) and polydopamine nanotubes.
研究领域:
分析化学
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分子生物学
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纳米技术
✓ 已验证
DOI
PubMed
高影响因子
JOURNAL OF HAZARDOUS MATERIALS
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2022
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IF 14.224
Jiamin Xu,Mingbin Liu,Weihua Zhao,Suqin Wang,Minfang Gui,Hongbo Li,Ruqin Yu
Lead ions are one of many common environmental pollutants, that can cause posing a serious threat to people’s health, thus, their efficient and sensitive detection is important. We propose a cascade signal amplification sensor using a DNAzyme-based strand displacement amplification (SDA) and hybridization chain reaction (HCR) for the high-sensitivity detection of Pb 2+ . In the demonstrated sensor system, the target metal ion can activate DNAzyme to cause a strand displacement reaction. Under the synergistic action of polymerase and nickase, large numbers of DNA strands are generated that can initiate HCR. The subsequent HCR can restore the fluorescence intensity of the FAM quenched for the fluorescence resonance energy transfer effect, which exhibits a strong fluorescence signal. The DNAzyme-based sensor allowed the detection of Pb 2+ down to 16.8 pM and resulted in a good dynamic line relationship ranging from 50 pM to 500 nM, and the biosensor also showed good selectivity . Furthermore, we confirmed that the proposed sensor can still detect lead ions in complex environments such as lake water, milk, and serum. We believe these findings will provide new ideas for the detection of toxic elements ions in the environment and food.
研究领域:
环境科学
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生物传感
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分子生物学
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DOI
PubMed
高影响因子
Chemical Biology & Drug Design
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2018
·Vol. 93
·期 6
·993-998
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IF 2.328
Ruimin Zang,Xin Wang,Yanyan Zhu,Tianming Yao,Shuo Shi
Exon 2 deletion of aminoacyl tRNA synthetase complex-interacting multifunctional protein 2 (AIMP2) is a genetic deletion related to various cancers, for instance ovarian and lung cancers. It can be worked as an indicator of cancer for diagnosis of diseases. Here, we developed a label-free method based on the formation of split G-quadruplex in the presence of target DNA combined with strand displacement to detect exon 2 deletion of AIMP2 (DE2) sensitively and selectively. This method is easy-operating and cost-saving. Moreover, it has observed discrimination of gene deletion from wild-types by naked eyes. The results demonstrate that this strategy can be further used for the detection of different gene deletions to achieve early diagnosis of diseases and allow better prognosis.
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
✓ 已验证
产品关联
DOI
PubMed
高影响因子
ANALYTICAL CHEMISTRY
·
2021
·
IF 6.986
Junhua Chen,Gu Shi,Chong Yan
A visual detection method was proposed for polychlorinated biphenyls (PCBs) detection using lateral flow test paper as the sensing platform. The aptamer sequence was used to recognize the target 3,3′,4,4′-tetrachlorobiphenyl (PCB77). The integration of Zn2+-dependent DNAzyme with toehold-mediated strand displacement reaction significantly improved the response signals. Gold nanoparticles were utilized as the signal tracers in the test paper, making the results visible directly by the naked eye. Under optimal conditions, the paper enables the visual detection of PCB77 as low as 10 pM without additional instrumentation. The assay displays a high selectivity for PCB77 against potential interfering molecules. The visual test paper is robust and has been applied to the detection of PCB77 in milk samples with good recovery and satisfactory accuracy. Using two different PCBs (PCB77 and PCB72) as inputs, we further fabricated OR and AND logic gates, which is conducive to the development of an intelligent detection strategy for PCBs monitoring. Given the attractive characteristics of disposability, low cost, logic operation, and intuitive output, the test paper shows great promise for on-site screening of PCBs in resource-limited areas.
研究领域:
分析化学
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环境科学
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生物传感
✓ 已验证
DOI
PubMed
高影响因子
ANALYTICAL CHEMISTRY
·
2020
·
IF 6.785
Xiaolun Peng,Huangli Yan,Zhen Wu,Wei Wen,Xiuhua Zhang,Shengfu Wang
Detection of biomarkers at the cellular level can provide more accurate and comprehensive information that is important for early diagnosis of diseases and evaluation of new drugs. However, the interference of a large number of components in cells and the requirement of high sensitivity bring great challenges for their detection. Herein, a robust and enzyme-free electrochemical platform was proposed for microRNA-21 (miRNA-21) detection by integrating the efficient separation of magnetic nanobeads (MBs) with the multisignal amplification of strand displacement amplification (SDA) and electrochemically mediated atom transfer radical polymerization (eATRP). The eATRP is capable of de novo growth of a number of electroactive polymers on the electrode surface for signal amplification. Compared to simple hybridization, SDA and eATRP can enhance the signals by ∼35-fold, achieving high signal-to-noise ratio for low-abundant target detection. Owing to their superparamagnetism and strong magnetic response ability, MBs endow the method with excellent specificity and anti-interference ability to detect miRNA-21 in cells. Using MBs as capture carriers, SDA and eATRP for signal amplification, and gold nanoflower (AuNF)-modified electrodes as working electrodes, as low as 0.32 aM miRNA-21 was detected. Furthermore, the successful detection of miRNA-21 in cells indicated the great prospect of this method in the early diagnosis of cancers, life science research, and single-entity electrochemical detection.
研究领域:
生物医学工程
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分子诊断学
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电化学传感器
✓ 已验证
DOI
高影响因子
MICROCHEMICAL JOURNAL
·
2021
·
IF 4.821
Qiao Xu,Ke Liu,Jingjing Jin,Xinfeng Zhang
The assay of biomarker proteins is of critical importance for disease diagnosis. In this work, we have designed a cascaded strand displacement reaction system for detection of low abundant biomarker protein in serum samples. The proposed binding-induced strand displacement conversion system, first converts a protein recognition event into the output of the DNA (cDNA) catalyst, and then the output cDNA catalytically load the reporter DNAzyme onto magnetic beads based on catalyzed hairpin assembly (CHA) reaction. The high density of the hairpin DNA on a magnetic bead surface results in an over 20-fold improvement in the reaction efficiency of CHA, compared with conventional CHA. In addition, the magnetic beads are able to efficiently separate the serum sample matrix that could remarkably affect the DNAzyme-catalyzed chemiluminescent (CL) reaction. This type of magnetic biosensor allows for the detection of 0.5 pM target protein in serum samples, whereas when using homogenous CHA biosensors, the CL signal for 2 nM PDGF-BB is seriously inhibited by the serum matrix. Since the sensing system integrates the protein recognition, signaling, signal amplification and matrix separation functions, it is promising for the assay of proteins in biological samples.
研究领域:
生物化学
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分子诊断
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纳米医学
✓ 已验证
DOI
高影响因子
Analytical Methods
·
2016
·Vol. 8
·期 39
·7054-7060
·
IF 1.915
Lili Yu,Hui Xu,Hou Chen,Liangjiu Bai,Wenxiang Wang
A label-free, exonuclease III (Exo III) assisted and cost-effective Hg2+ fluorometric assay based on a strand displacement amplification strategy was developed with high sensitivity and selectivity. Thymine–thymine (T–T) mismatches between the toehold domains in hairpin DNA and assistant DNA can serve as specific recognition elements for Hg2+ binding with the help of T–Hg2+–T base pairs to initiate a toehold-mediated strand displacement reaction. Exo III provides a reduced background signal, and the method is label-free using an intercalator, Sybr green I (SG). The number of T–T mismatches, the concentration of SG and Exo III, the incubation temperature and time were all optimized. Under the optimized condition, the limit of detection for Hg2+ can be as low as 9.42 pM with a linear range from 0.01 to 1 nM. Moreover, this proposed strategy shows excellent selectivity, and was successfully applied for the detection of Hg2+ in laboratory tap water and Yanjing lake water samples in our university.
研究领域:
生物化学
·
环境科学
✓ 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）