ANALYTICA CHIMICA ACTA
·
2025
·
IF 6
Fei Ding,Xiaomin Hang,Sen Tian,Wei Cao,Jingwen Wu,Li Wang
Primer exchange reaction (PER) is a novel and simple nucleic acid-templated extension technique that has recently attracted much attention in the field of biosensing. However, current PER reactions have shown relatively slow rates and low amplification performances, resulting in long assay times and limited detection sensitivities. Here we report a nicking endonuclease-mediated PER reaction (named NEPER) that rapidly releases amplified DNA products by adding a nicking endonuclease to hydrolyze the hybridized double-stranded DNA (dsDNA), and consequently has a maximum speed that is thirty orders of magnitude greater than the maximum for conventional PER. We further combined a CRISPR/Cas12a signal readout technique and developed a cascade NEPER-CRISPR/Cas12a method that can detect miRNA-155 with a limit of detection (LOD) down to 3.1 fM. We also show that the NEPER-CRISPR/Cas12a can be used to detect targets in serum samples.
研究领域:
分子生物学
·
生物传感
·
诊断学
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产品关联
DOI
PubMed
高影响因子
Chemosensors
·
2022
·Vol. 10
·期 7
·276
·
IF 4.229
Kunhan Nie,Yongjian Jiang,Na Wang,Yajun Wang,Di Li,Lei Zhan,Chengzhi Huang,Chunmei Li
The abnormal expression of miRNA is closely related to the occurrence of pancreatic cancer. Herein, a programmable DNAzyme amplifier for the universal detection of pancreatic cancer-related miRNAs was proposed based on its programmability through the rational design of sequences. The fluorescence signal recovery of the DNAzyme amplifier showed a good linear relationship with the concentration of miR-10b in the range of 10–60 nM, with a detection limit of 893 pM. At the same time, this method displayed a high selectivity for miR-10b, with a remarkable discrimination of a single nucleotide difference. Furthermore, this method was also successfully used to detect miR-21 in the range of 10–60 nM based on the programmability of the DNA amplifier, exhibiting the universal application feasibility of this design. Overall, the proposed programmable DNAzyme cycle amplifier strategy shows promising potential for the simple, rapid, and universal detection of pancreatic cancer-related miRNAs, which is significant for improving the accuracy of pancreatic cancer diagnosis.Keywords:DNAzyme amplifier;signal amplification;miRNA detection;pancreatic cancer
研究领域:
分子生物学
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医学诊断
·
生物化学
✓ 已验证
产品关联
DOI
高影响因子
APPLIED BIOCHEMISTRY AND BIOTECHNOLOGY
·
2024
·
IF 3.1
Yan Na,Hu Zhiwei,Zhang Lingling
The development of a bio-sensing strategy based on CRISPR/Cas that is exceptionally sensitive is crucial for the identification of trace molecules. Colorimetric miRNA detection utilizing CRISPR/Cas13a-triggered DNAzyme signal amplification was described in this article. The developed strategy was implemented for miRNA-21 detection as a proof of concept. The cleavage activity of Cas13a was triggered when the target molecule bonded to the Cas13a-crRNA complex and cleaved uracil ribonucleotides (rU) in the substrate probe. As a consequence, the S chain was liberated from the T chain that had been modified on magnetic beads (MB). The G-rich sections were then exposed when the catalytic hairpin assembly between the H1 and H2 probes was activated by the released T@MB. G-rich section can fold into G-quadruplex. By catalyzing the formation of green ABTS 3 – via HRP-mimicking G-quadruplex/hemin complexes, colorimetric measurements of miRNA can be achieved visually through DNAzyme-mediated signal amplification. The method demonstrated a low limit of detection of 27 fM and a high selectivity towards target miRNA eventually. As a result, the developed strategy provides a clinical application platform for the detection of miRNAs that is both ultrasensitive and extremely specific.
研究领域:
生物技术
·
分子生物学
·
诊断方法
✓ 已验证
DOI
PubMed
高影响因子
BIOELECTROCHEMISTRY
·
2022
·
IF 5.76
Qian Liu,Huawei Shen,Baiying Li,Juan Cai,Yang Peng,Zhi Weng,Hongyan Yu,Guoming Xie,Wenli Feng
Currently, developing an effective and easy-to-operate signal amplification assay to detect the trace-amount miRNAs in serum remains a significant challenge. Herein, an ultrasensitive CeO 2 @Ag hybrid nanoflower (CeO 2 @Ag HNF)-labeled electrochemical biosensor was developed for sensing miRNA, based on a target-feedback rolling-cleavage (TFRC) signal amplifier. CeO 2 @Ag HNFs possessing a unique three-dimensional layered structure were synthesized without any complex reaction conditions, such as heating and vacuum. The bimetallic nanoflowers provided a large surface area and excellent CAT-like activity, thereby enhancing electrochemical performance. Based on the principle of branched catalytic hairpin assembly, target miRNA could continuously trigger the assembly of branched junctions with ends composed of DNAzyme. The activated DNAzyme was able to cleave the hairpin substrate efficiently and release to capture more CeO 2 @Ag HNFs label. The process combining target-driven branched catalytic hairpin assembly and DNAzyme-assisted rolling cleavage were defined as TFRC signal amplification. This sensitive electrochemical biosensor exhibited good linear relationship of 1 fM − 1 nM with a detection limit down to 0.89 fM. The proposed method is expected to have a promising application in the miRNA-associated fundamental research and clinical diagnosis.
研究领域:
生物化学
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分子生物学
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纳米医学
✓ 已验证
DOI
高影响因子
INTERNATIONAL JOURNAL OF BIOLOGICAL MACROMOLECULES
·
2025
·
IF 8.5
Hui Li,Zongshan Zhang,Wantong Hao,Huixin Shi,Ruiyang Yin,Jing Xu
MicroRNAs (miRNAs) are key regulators of gene expression and play a crucial role in various biological processes. However, existing methods for miRNA detection face challenges in terms of sensitivity, specificity and ease of use. These limitations highlight the need for innovative and highly sensitive detection platforms capable of accurately quantifying miRNAs. In this work, an integrated biochip based on a self-powered sensor system developed for the ultra-sensitive detection of miRNAs-21 and miRNA-20a. The system uses a bilirubin oxidase (BOD)-supported bio-cathode and a glucose oxidase (GOD)-supported bio-anode to form a closed loop that converts chemical energy into electrical energy. The complementary chains of target miRNAs are modified on the bio-anode using a bio-conjugate amplification strategy, enabling the system to generate an electrical signal proportional to the concentration of miRNAs. The sensor has a wide linear range of 5 fmol/L to 10 nmol/L for miRNA-20a and 1 fmol/L to 10 fmol/L for miRNA-21, with low detection limits of 0.176 fmol/L and 0.138 fmol/L, respectively. These detection limits are significantly lower than those achieved by existing technologies, making our biosensor a significant advancement in the field. In addition, the sensor system is integrated into a portable chip that transmits electrical signals in real time via Bluetooth to a mobile phone interface, enabling convenient and rapid analysis.
研究领域:
生物医学工程
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分子生物学
·
分析化学
✓ 已验证
产品关联
DOI
PubMed
高影响因子
ANALYTICAL CHEMISTRY
·
2019
·
IF 6.35
Jianguo Xu,Chao Yan,Xinxin Wang,Bangben Yao,Jianfeng Lu,Guodong Liu,Wei Chen
Facile and efficient assembly of intelligent DNA nano-objects with the ability to exert robust microRNA determination is essential for DNA nanobiotechnology and basic biomedical study. Herein, we present a novel target-triggered DNA assembly pathway for the construction of multibranched DNA nanoarchitectures by the combination of DNA concatamers with G-quadruplex wires. The DNA concatamers acting as the main body structures were assembled by hybridization chain reaction (HCR), while the G-quadruplex wires acting as the branch structures were generated by terminal deoxynucleotidyl transferase (TdT)-promoted polymerization and G-quadruplex units (GUs) based self-assembly. With a copious number of G-quadruplex replicates collected on the electrode surface, the obtained multibranched DNA nanoarchitectures were able to bind with lots of hemin, which further led to a significantly amplified electrochemical sensing signal for ultrasensitive and selective detection of miRNA-21. The detection limit was achieved as low as 0.2 fM with a wide dynamic response ranging from 10 fM to 100 nM, which is much more powerful or at least comparable to most of the reported studies. Furthermore, the electrochemical biosensor offered an excellent specificity, and even the single-base mutation against the perfectly matched target miRNA can be easily distinguished easily. Furthermore, the real biological samples were also desirably analyzed, indicating that the proposed strategy is an ideal platform for the fabrication of versatile DNA nanobiosensors.
研究领域:
DNA纳米生物技术
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生物医学研究
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分子诊断
✓ 已验证
产品关联
DOI
PubMed
高影响因子
BIOSENSORS & BIOELECTRONICS
·
2025
·
IF 10.5
Wenrui Xu,Xiaoguang Gao,Yuyan Zhai,Xiaoliang Zhang,Yingqian Wang,Rong Deng,Xiaochun Li
It remains a challenge to achieve high-sensitivity detection of tumor marker miRNA using optical refractive index (RI) sensors without nucleic acid amplification. This study proposes the photothermal effect-assisted reduced graphene oxide (rGO) biosensor that combines the photothermal effect of rGO with the rGO-based RI sensor for high-sensitivity detection of tumor marker miRNA-21. The rGO was functionalized with DNA probes capable of specifically hybridizing with the target miRNA-21. Quantitative detection of miRNA-21 was achieved by monitoring the RI change caused by the competitive hybridization of single-strand DNA (ssDNA)-functionalized AuNPs and target miRNA-21 with the DNA probes on the rGO surface. The presence of AuNPs disturbed the evanescent field on the rGO surface, thus achieving signal amplification. Furthermore, the localized photothermal effect heat induced by the interaction between rGO and pump light can effectively improve the hybridization kinetics of nucleic acid chains and achieve further signal amplification. The proposed biosensor had a high sensitivity toward the target miRNA-21, achieving a low detection limit of 4.05 fM without nucleic acid amplification. Its high specificity allowed for the recognition of single-base mismatches in miRNA-21. In addition, accurate quantification of low abundance miRNA-21 spiked into human urine samples was also successfully achieved. The photothermal effect-assisted rGO biosensor offers a promising approach for high-sensitivity detection of tumor marker miRNA without need for nucleic acid amplification.
研究领域:
生物传感器技术
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分子诊断学
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肿瘤标志物研究
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产品关联
DOI
PubMed
高影响因子
LAB ON A CHIP
·
2019
·Vol. 19
·期 10
·1783-1789
·
IF 6.914
Lijun Cai,Feika Bian,Lingyu Sun,Huan Wang,Yuanjin Zhao
The ultrasensitive detection of multiple nucleic acids has great significance in a broad range of medical fields since nucleic acid samples are usually of small volumes and highly diluted. Herein, we present a novel magnetic photonic crystal (PhC) barcode-integrated condensing-enriched superhydrophobic platform for the ultrasensitive multiple miRNA detection. Droplets containing targets could suck the hydrogel PhC barcodes without losing their targets due to the difference in wettability between the hydrophilic PhC barcodes and hydrophobic substrate. During the evaporation of water from the droplet microreactors, these targets could be effectively enriched in the barcodes for achieving higher sensitivities. As the encoding signals of the PhC barcodes are the characteristic reflection peaks generated by their ordered physical nanostructures, the barcodes are stable and free from any fluorescent background and photobleaching; thus, they are accurate for distinguishing different targets. In addition, with the integration of magnetic nanoparticles into the hydrogel PhC barcodes, they could be imparted with the features of immobilization during change of medium and flexible movement for controlling the reaction, both of which could facilitate the detection processes. Based on this platform, we have demonstrated that the detection limit of multiple miRNAs is improved by about three orders. Thus, our platform is an ideal alternative for the ultrasensitive simultaneous multiplex analysis in medical fields.
研究领域:
生物医学工程
·
分子诊断学
✓ 已验证
产品关联
DOI
PubMed
高影响因子
TALANTA
·
2021
·
IF 6.057
Yunpeng Han,Rong Zou,Lingyun Wang,Chunyan Chen,Hang Gong,Changqun Cai
Herein, a metal-organic framework (UiO-66-NH 2 ) with two functions (intrinsic fluorescence and fluorescence quenching ability) is designed to establish a ratiometric fluorescent platform for high-performance miRNA detection. The use of a fluorescent organic ligand endows the MOF material with a strong intrinsic fluorescence at 440 nm. In the presence of target miRNA, the fluorescence signal of the FAM is restored with the triple helix molecular beacons bind to the target. Using the I FAM /I MOF signal as the output, the prepared ratiometric probe was able to eliminate disturbance caused by the sensing environment. Under the optimal reaction conditions, including buffer pH of 7.4, temperature of 37 °C, and response time of 1 h, the best detection results can be obtained. The ratiometric fluorescence probe presented showed good sensitivity and selectivity for detecting miRNA-203 and the limit of detection was 400 pM with a wide linear range from 1 nM to 160 nM. In addition, this method was applied to diluted human serum and cell lysates, and good detection effect was realized.
研究领域:
生物化学
·
分子生物学
·
分析化学
✓ 已验证
产品关联
DOI
PubMed
高影响因子
Analytical Methods
·
2022
·Vol. 14
·期 44
·4523-4530
·
IF 3.532
Jiao Wang,Liyuan Wen,Ruyuan Cao,Xiaorong Gao,Xia Li,Ensheng Xu,Qi Zhang,Shuling Xu,Caifeng Dai,Qingwang Xue
Single miRNAs are not specific and accurate enough to meet the strict diagnosis requirements in practice. Therefore, simultaneous monitoring of multiplexed miRNA in biological samples can not only improve the accuracy and specificity of bioassays but also avoid the squandering of valuable biological specimens. Herein, we designed a metal sulfide nanoparticle-based dual barcode-triggered DNAzyme cascade strategy for the sensitive and simultaneous multiplex miRNA detection in a single assay. Firstly, the capture probes (H1, H2) specifically recognize targets (miRNA-21, miRNA-141), exposing the stem of H1 and H2. Then, with the introduction of a detection probe (CuS-H3, ZnS-H4), the exposed H1 and H2 catalyze the hairpin assembly (CHA) reaction, realizing target miRNA recycling, and forming H1/H3-CuS and H2/H4-ZnS complexes. Subsequently, the formed H1/H3-CuS and H2/H4-ZnS complexes are encoded on magnetic beads through the biotin/streptavidin interaction. The CuS and ZnS nanoparticles captured by magnetic beads release thousands of Cu2+ and Zn2+via the cation exchange reaction. Finally, the released Cu2+ and Zn2+ specially activate the DNAzyme of the catalytic and molecular beacon (CAMB) system. The CAMB system affords an amplified fluorescence signal output by cycling and regenerating the metal ion-dependent DNAzyme to realize multiple enzymatic turnovers. Benefiting from target recycling, nanoparticle amplification, and catalytic and molecular beacon amplification, there is substantial amplification and the target miRNAs can be detected at 0.06 fM (miRNA-21) and 0.048 fM (miRNA-141) in a single assay. Furthermore, the high selectivity and accuracy of the assay were proved by practical analysis of different cancer cells, which exhibited good practicability in multiplex miRNA detection in clinical sera. The results indicate that the proposed strategy holds great potential for the sensitive detection of multiplex cancer biomarkers and offers the opportunity for future applications in clinical diagnosis.
研究领域:
生物化学
·
分子诊断
·
纳米医学
✓ 已验证
产品关联
DOI
PubMed
高影响因子
ELECTROANALYSIS
·
2022
·Vol. 35
·期 2
·e202200190
·
IF 3.077
Qi Zhang,Zhihui Guo,Xingwang Zheng
Polyethyleneimine (PEI) is a cationic polymer with a lot of amino groups in its molecular structure that is secure, sterile, and biocompatible. However, it is difficult for PEI molecules to be directly applied in electroanalysis due to their poor electroactivity. Herein, the PEI-based carbonized polymer nanodots (CPDs) with core-shell structure were prepared by hydrothermal method using PEI as the precursors. The results showed that the as-prepared PEI-based CPDs can not only exhibit the unique properties of carbon nanodots but also inherit the properties of PEI. Subsequently, based on the strong coordination interaction between the abundant amino groups on the surface of PEI and silver ions, the electroactive Ag@carbonized polymer dots (Ag@CPDs) were prepared. Furthermore, to investigate the electrochemical behaviors and application of Ag@CPDs, the DNA molecule (double-stranded and single-stranded DNA) was taken as an example. It was found that the interaction of DNA with Ag@CPDs had a significant influence on the current intensity of Ag@CPDs. Based on this phenomenon, a sensitive electrochemical technique for miRNA detection was established and effectively employed to detect miRNA in human serum samples based on the discriminating effect of Ag@CPDs towards ssDNA probes and ssDNA probe/miRNA complex (dsDNA).
研究领域:
材料科学
·
生物医学工程
·
分析化学
✓ 已验证
产品关联
DOI
高影响因子
ANALYTICA CHIMICA ACTA
·
2023
·
IF 6.2
Yang Lu,Yiyue Yu,Yeru Wang,Wenbo Zhou,Ziyi Cheng,Liandong Yu,Shaojiang Zheng,Rongke Gao
Background Precise and specific miRNA detection plays a vital role in exploring development mechanisms of cancer disease, thereby it can significantly improve relevant prevention and treatment strategies. Results In this work, a surface-enhanced Raman spectroscopy (SERS)-based microfluidic chip has been devised with a microcone array SERS substrate (MCASS) for the miR-141 detection. This substrate excels in unique SERS activity and large surface area for DNA oligonucleotide modification. As the presence of miR-141, the DNAzyme walker induced cleavage reaction took place on the finely designed and prepared dual DNA conjugated SERS nanoprobes. The SERS nanoprobes can anchor on MCASS by the DNA hybridization that achieved an impressive detection limit in the femtomolar level. Significance With this integrated SERS-based microfluidic chip, we provided a miRNA detection strategy using DNAzyme walker amplification technology. It is believed that this strategy could be a powerful tool for miRNA detection and related cancer screening test .
研究领域:
生物医学工程
·
分子生物学
·
纳米技术
✓ 已验证
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）