LUMINESCENCE
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2020
·Vol. 35
·期 6
·835-844
·
IF 1.855
Zheng Chang,Jing Feng,Xingwang Zheng
In this paper, a convenient reverse-phase microemulsion method for the synthesis of SiO 2 nanoparticles (NPs) by simply introducing the chitosan and fluorescent dye of lucigenin during the formation reaction of SiO 2 NPs was proposed. Addition of chitosan can make the SiO 2 NPs porous, and increases lucigenin molecule incorporation into chitosan/SiO 2 NPs nanopores based on electrostatic interaction and supermolecular forces. Therefore, fluorescence quantum yield of the lucigenin/chitosan/SiO 2 composite nanoparticles was increased by introduction of chitosan and compared with lucigenin/SiO 2 NPs without chitosan. Because the number of negative charges carried when using single-stranded DNA (ssDNA) was different from that of double-stranded DNA (dsDNA), the numbers of lucigenin/chitosan/SiO 2 composite nanoparticles with positive charge adsorbed using ssDNA or dsDNA were different. Consequently, fluorescence intensity caused using ssDNA or dsDNA/miRNA was clearly discriminative. With increase in target DNA/miRNA concentration, the difference in fluorescence intensity also increased, resulting in a good linear relationship between fluorescence intensity sensitizing value and target miRNA concentrations. Therefore, a new fluorescence analysis method for direct detection of let-7a in human gastric cancer cell samples without enzyme, label free and no immobilization was established using lucigenin/chitosan/SiO 2 composite nanoparticles as a DNA hybrid indicator. The proposed method had high sensitivity and selectivity, low cost and the detection limit was 10 fM (S/ N = 3).
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高影响因子
JOURNAL OF NANOBIOTECHNOLOGY
·
2025
·
IF 12.6
Huang Xiaobin,Yi Yakun,Zhou Ke,Liu Lei,Wu Hai-Chen
MicroRNAs (miRNAs) play crucial roles in the pathogenesis of diverse diseases; however, their extremely low abundance in complex biological systems makes highly sensitive and specific multiplexed detection particularly challenging. Here, we report an enzyme-assisted nanopore sensor that enables simultaneous and ultrasensitive detection of multiple miRNAs. Capture DNA (cDNA) immobilized on streptavidin-coated magnetic beads specifically hybridizes with target miRNAs, opening the cDNA hairpin and facilitating the binding of alkaline phosphatase (ALP)-labeled detection DNA to form stable complexes. ALP catalyzes the dephosphorylation of designed phosphorylated peptide probes, inducing conformational and charge changes that yield distinct current blockade signatures in the presence of cucurbit [ 7 ]uril within the nanopore. This strategy reduces background interference via magnetic separation and amplifies detection signals through enzymatic reactions, thereby markedly improving sensitivity for low-abundance miRNAs. The sensor achieves a broad dynamic range from 10.0 fM to 10.0 nM across three target miRNAs, with femtomolar-level limits of detection. By integrating molecular separation, enzymatic amplification, and nanopore sensing, this platform provides a robust approach for precise miRNA analysis, offering strong potential for early disease diagnosis.
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TALANTA
·
2021
·
IF 6.057
Chunrong Li,Yanfei Zhang,Jun Chen,Liping Liu,Huihui Yang,Si-Yang Liu,Yuzhi Xu,Zong Dai,Xiaoyong Zou
Nanomaterial-based on-off-on fluorescence sensing strategies are significant particularly in intracellular nucleic acids imaging assay. There still remains challenge to rationally balance fluorescence quenching efficiency and recovery dynamics. We assume that the performance of on-off-on fluorescence sensing strategy can be fundamentally improved on small zero-dimensional (0D) nanomaterial with precisely modulated surface charge. For a proof-of-concept demonstration, silicon nanoparticle (SiNP) with ~4 nm was synthesized and used as the quencher model, of which the surface charge density was modulated by modification of triphenylphosphonium (TPP). The influence of particle size, surface charge and charge density of the nanomaterials on sensing performance was systematically investigated. The strategy showed a low limit of detection (LOD) as 26 pM for target model miR-494, which is one of the lowest in nanomaterial-based on-off-on sensing platforms. And the LOD is even comparable to amplification-based methods in a greatly shortened assay time (2.5 h). The miR-494 expresses in cancerous and normal living cells of human cervical carcinoma (HeLa), human lung carcinoma (A549), human breast cancer (MCF-7), and normal human mammary epithelial (MCF-10A) cells were imaged and localized with significantly improved sensitivity and specificity. These excellent performances insure it a promising candidate as convenient and non-enzymatic sensing platform for miRNA-associated disease detection and early diagnosis.
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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高影响因子
TALANTA
·
2017
·
IF 4.162
Shiyu Li,Kui He,Rong Liao,Chunyan Chen,Xiaoming Chen,Changqun Cai
An interference-free and label-free sensing platform was developed for the highly sensitive detection of microRNA-21 (miRNA-21) in vitro by magnetic silicon microsphere (MNP)-reduced graphene oxide (rGO)-based sandwich probe. In this method, DNA capture probes (P 1 ) were connected with MNPs at the 5′ end and hybridized with completely complementary target miRNA. Subsequently, rGO was retained and induced the fluorescence quenching in the supernatant. Through the magnetic separation, the supernatant environment was simplified and the interference to analytical signal was eliminated. When DNA capture probe-modified magnetic silicon microspheres (MNP-P 1 ) were adsorbed through rGO in the absence of a target and formed a sandwich structure, the formed nanostructure was easily removed from the solution by a magnetic field and the fluorescence intensity was maximally recovered. This proposed strategy, which both overcame the expensive and cumbersome fluorescent labeling , and eliminated interference to analytical signal for guaranteeing high signal-to-background ratio, exhibited high sensitivity with a detection limit as low as 0.098 nM and special selectivity toward miRNA-21. The method was potentially applicable for not only detection of miRNA-21 but also various biomarker analyses just by changing capture probes.
研究领域:
生物传感器
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纳米技术
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分子诊断
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分析化学
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高影响因子
TALANTA
·
2022
·
IF 6.556
Qiujun Lu,Lingling Xie,Shuhang Yin,Feng Chen,Cuiyan Wu,Meiling Liu,Haitao Li,Youyu Zhang
The overexpression of microRNA-10b (miRNA-10b) is closely associated with hepatocellular carcinoma (HCC), which may act as an advance biomarker to diagnose HCC early. Here, a novel luminescent nanosensor for the ultrasensitive detection of miRNA-10b was firstly developed by target-triggered catalytic hairpin assembly ( CHA ) and upconversion nanoparticles (UCNPs)-based luminescence resonance energy transfer (LRET). The hybridization process of DNA1-modified UCNPs with DNA2-modified gold nanoparticles (AuNPs) makes the energy donor (UCNPs) and acceptor (AuNPs) into close proximity, resulting in luminescence quenching of UCNPs. The miRNA-10b can hybridize with the hairpin structure of DNA2, releasing the recognition domain site and activate the CHA reaction with its signal amplification chain (H3). At the same time, the LRET was inhibited accompany with the AuNPs moved away from the UCNPs, which resulted in the luminescence recovery of UCNPs. The miRNA-10b can be quantified by measurement of the luminescence recovery rate of UCNPs. The sensor has high sensitivity with a detection limit of 8.96 fM. In addition, the sensor can be used to the discrimination of miRNA-10b expression levels in human hepatocarcinoma and hepatocyte cell lines with satisfactory results. The technique shows great promise for early diagnosis of HCC because of its high sensitivity and selectivity .
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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高影响因子
ANALYTICAL CHEMISTRY
·
2017
·
IF 6.32
Xiangmin Miao,Zhiyuan Cheng,Haiyan Ma,Zongbing Li,Ning Xue,Po Wang
A novel strategy was developed for microRNA-155 (miRNA-155) detection based on the fluorescence quenching of positively charged gold nanoparticles [(+)AuNPs] to Ag nanoclusters (AgNCs). In the designed system, DNA-stabilized Ag nanoclusters (DNA/AgNCs) were introduced as fluorescent probes, and DNA–RNA heteroduplexes were formed upon the addition of target miRNA-155. Meanwhile, the (+)AuNPs could be electrostatically adsorbed on the negatively charged single-stranded DNA (ssDNA) or DNA–RNA heteroduplexes to quench the fluorescence signal. In the presence of duplex-specific nuclease (DSN), DNA–RNA heteroduplexes became a substrate for the enzymatic hydrolysis of the DNA strand to yield a fluorescence signal due to the diffusion of AgNCs away from (+)AuNPs. Under the optimal conditions, (+)AuNPs displayed very high quenching efficiency to AgNCs, which paved the way for ultrasensitive detection with a low detection limit of 33.4 fM. In particular, the present strategy demonstrated excellent specificity and selectivity toward the detection of target miRNA against control miRNAs, including mutated miRNA-155, miRNA-21, miRNA-141, let-7a, and miRNA-182. Moreover, the practical application value of the system was confirmed by the evaluation of the expression levels of miRNA-155 in clinical serum samples with satisfactory results, suggesting that the proposed sensing platform is promising for applications in disease diagnosis as well as the fundamental research of biochemistry.
研究领域:
生物化学
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分子诊断
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纳米生物学
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高影响因子
MICROCHEMICAL JOURNAL
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2026
·
IF 5.2
Yaru Zhang,Yongchang Sun,Jing Yuan,Lihua Ding,Leiliang He,Yongjun Wu,Songcheng Yu
MicroRNAs (miRNAs), as crucial regulators of gene expression, play significant roles in cancer development. Among them, miRNA-21 has been recognized as a promising biomarker for the diagnosis and prognosis evaluation of multiple cancers. However, existing CRISPR/Cas12a-based miRNA detection methods generally require reverse transcription and pre-amplification steps, which complicate the operational workflow and are prone to introducing cross contamination. Furthermore, the dependence on single-fluorescence signal detection makes them vulnerable to environmental and instrumental fluctuations, which results in risk of false-positive outcomes. Herein, we developed a ratiometric and sensitive CRISPR/Cas12a system with trans-cleavage-triggered feedback signal amplification for amplification-free detection of miRNA-21. The strategy employs a novel multifunctional switch probe that integrates target recognition, signal transduction, and cascade amplification within a unified system. A trans-cleavage-triggered feedback signal amplification circuit was constructed to enhance detection sensitivity without the need for additional enzyme systems. Moreover, a ratiometric fluorescence signal was implemented by calculating the fluorescence intensity ratio between CY5 and Auramine O, which effectively minimizes interference from environmental and instrumental variations and substantially improves detection accuracy and reliability. Experimental results demonstrate that the method achieves a detection limit of 1.28 nM, along with excellent specificity, accuracy, and reproducibility. In real sample analysis, differential expression levels of miRNA-21 in lung adenocarcinoma cells (A549) and non-cancerous adult bronchial epithelial cells (BEAS-2B) was observed, which highlights its potential value for clinical diagnostics.
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高影响因子
SENSORS AND ACTUATORS B-CHEMICAL
·
2025
·
IF 7.7
Xin Xie,Peng Chen,Jingxi Wang,Ding Wang,Daxiu Li,Beibei Kou,Wenjiao Zhou
Herein, a novel electrochemical biosensor on the basis of MB/AuNPs@Zr-MOF signal probe and enzyme-free cascade signal amplification was fabricated for the ultrasensitive detection of colorectal cancer (CRC)-associated miR-92a-3p. Impressively, compared with the traditional electrochemical probes, MB/AuNPs@Zr-MOF not only exhibits high loading for electroactive substances, but also shows enhanced electronic conductivity, thereby generating stable and high electrochemical signal. Meanwhile, under the assistance of the target-induced enzyme-free cascade signal amplification including multicomponent nuclease (MNAzyme) recycle catalytic and catalytic hairpin assembly (CHA), the sensitivity of proposed biosensor was further improved. Additionally, the DNA tetrahedral scaffold (TDNs) was served as an electrode support interface, which could greatly reduce the non-specific adsorption and improve the analytical performance. As a result, the developed biosensor enabled the miR-92a-3p with a detection limit as low as 0.36 fM. This work provided an efficient method to increase the sensitivity and the stability of electrochemical sensing platforms, showing great potential for the detection of biomarker and disease diagnosis.
研究领域:
生物传感器
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纳米技术
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电化学
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癌症诊断
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分子生物学
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生物分析化学
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高影响因子
MICROCHEMICAL JOURNAL
·
2025
·
IF 5.1
Guobin Huang,Weiting Xu,Jinxin Chi,Chenchen Lin,Jiabin Wang,Xucong Lin
Environmental smog pollution induces chronic obstructive pulmonary disease (COPD), and causes the urgent need of timely, safe and quantitative testing for risk assessment. The miRNA-based bioanalysis offers a non-invasive access to rapid COPD screening, while challenges remain in accurately identifying miRNAs from complex matrices. Herein, a novel bionic aptasensor has been pioneered for meeting the specific and sensitive detection of the miR-155 in serum for COPD diagnosis, integrating tetrahedral framework nucleic acid (tFNA) confined affinity recognition and enzyme-free hybridization chain reaction (HCR) amplification strategy. Precision-designed tFNAs were fabricated and hybridized with FAM-labeled cDNAs, enabling dual-mode recognition mechanism including spatially confined aptamer recognition and bionic molecular sieve size-selective action. Proteins and pre-miRNAs were excluded, and the target microRNA-155 was specifically captured. This triggered a structural switch of tFNAs and activated cDNA-mediated HCR, assembling double-stranded DNA and achieving exponential fluorescence enhancement synergistically produced by SYBR Green I (SGI) and FAM. The optimal limit of detection as 53 fM for miRNA-155 was achieved with laser-induced fluorescence (LIF). Applied to COPD diagnosis, the detections of trace miRNA-155 were satisfactory and the recovery yields were ranged in 96.4 ± 5.2 % ∼106.2 ± 2.1 % ( n = 3) in the serum of COPD patients. Smoke-exposed model rats were also analyzed, and the miRNA-155 was achieved with a significant concentration up-regulation to light the COPD. It offers a high-performance protocol for meeting the non-invasive rapid screening of COPD, further providing promising technologies for evaluating safety issues of environmentally induced diseases.
研究领域:
生物传感器
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分子诊断学
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环境健康
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RNA生物学
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肺部疾病研究
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高影响因子
INTERNATIONAL JOURNAL OF BIOLOGICAL MACROMOLECULES
·
2024
·
IF 7.7
Mengxin Zhang,Yingyu Zhang,Qionglin Wang,Kangbo Liu,Lifeng Li,Zhidan Yu,Ligong Hou,Xianwei Zhang,Huan Zhao,Weyland Cheng,Wancun Zhang
In situ imaging of microRNA (miRNA) content and distribution is valuable for monitoring tumor progression. However, tumor specific in situ imaging remains a challenge due to low miRNA abundance, lack of biological compatibility, and poor specificity. In this study, we designed a DNA tetrahedral framework complex with hairpins (DTF-HP AP ) consisting of an apurinic/apyrimidinic site (AP site) that could be specifically recognized and cleaved by apurinic/apyrimidinic endonuclease 1 (APE1). Efficient and specific in situ imaging of miR-21 in tumors was thus achieved through catalytic hairpin assembly (CHA) reaction. In this study, DTF-HP AP was successfully constructed to trigger the cumulative amplification of fluorescence signal in situ . The specificity, sensitivity and serum stability of DTF-HP AP were verified in vitro , and DTF-HP AP could be easily taken up by cells, acting as a biosensor to detect tumors in mice. Furthermore, we verified the ability of DTF-HP AP to specifically image miR-21 in tumors, and demonstrated its capability for tumor-specific imaging in clinical samples.
研究领域:
分子生物学
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生物医学工程
·
肿瘤学
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高影响因子
MICROCHIMICA ACTA
·
2020
·Vol. 187
·期 4
·1-9
·
IF 6.232
Cao Xiaodong,Zhang Kairui,Yan Wuwen,Xia Zihao,He Shudong,Xu Xuan,Ye Yongkang,Wei Zhaojun,Liu Songqin
A selective and sensitive fluorescence biosensor is described for determination of microRNA-167 using fluorescent resonant energy transfer (FRET) strategy. The FRET system comprises carbon dots (CDs, donor) labeled with probe DNA (pDNA) and polydopamine (PDA)-coated Fe 3 O 4 nanoparticles (Fe 3 O 4 @PDA NPs, acceptor). The CDs-pDNA can be absorbed onto the surface of Fe 3 O 4 @PDA NPs because of the strong π interaction between pDNA and PDA. With the enhanced adsorption ability of Fe 3 O 4 @PDA NPs by Ca 2+ , the fluorescence intensity of CDs at 445 nm (excitation at 360 nm) is quenched. In presence of microRNA-167, the hybridized complex of CDs-pDNA-microRNA-167 will be released from the surface of Fe 3 O 4 @PDA NPs due to the weak π interaction of the complex and PDA. This results in the fluorescence recovery of CDs. By application of twice-magnetic separation, the biosensor shows a wide linear range of 0.5–100 nM to microRNA-167 with a 76 pM detection limit. The method was applied to the determination of microRNA-167 in samples of total microRNA extractions from A. thaliana seedlings, and the recoveries ranged from 96.4 to 98.3%.
研究领域:
生物化学
·
分子生物学
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高影响因子
ANALYTICAL CHEMISTRY
·
2021
·
IF 6.986
Jiao Yang,Yanzhou Li,Longhua Guo,Bin Qiu,Zhenyu Lin
Conventional photosensitive materials such as TiO2 suffer from restricted absorption in the ultraviolet region, fast recombination of photogenerated electron–hole pairs, and a lack of functional groups for biocoupling, which hinder their application in photoelectrochemical (PEC) biosensing. Herein, a new coordination polymer (CP) based on Cu(I), chloridion, and 4-mercaptobenzoic acid (MBA) has been designed and synthesized (called CuClx(MBA)y). The prepared p-type CuClx(MBA)y exhibits visible-light absorption due to its narrow optical band gap (2.59 eV), and its proper band edge position enables it to form a p–n junction with TiO2. Through layer-by-layer assembling, the photocurrent intensity of the CuClx(MBA)y/TiO2/FTO composite photoelectrode was 3.7-fold higher than that of a TiO2/FTO electrode and 35-fold higher than a CuClx(MBA)y/FTO electrode. The potential enhancement mechanism was discussed, which lies in the contributions of CuClx(MBA)y in enhancing absorption in the visible-light region and boosting the separation of electron–hole pairs of TiO2 by the p–n junction. Furthermore, CuClx(MBA)y nanosheets can realize bioconjugation directly, thanks to its abundant carboxyl groups. The CuClx(MBA)y/TiO2/FTO composite photoelectrodes were applied to develop a sensitive PEC biosensor for microRNA-21 (model target). By subtly exploiting the energy transfer between CuClx(MBA)y and Au nanoparticles (AuNPs), AuNPs served as effective quenchers. In the presence of the target, AuNP-labeled sDNA1 connected to the electrode surface, and thus, a decreased photocurrent was obtained. The proposed biosensor has a low detection limit of 0.29 fM (S/N = 3), good selectivity, and reproducibility. The proposed system was applied to monitor microRNA in cancer cells with satisfying results.
研究领域:
生物化学
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分析化学
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纳米技术
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分子生物学
·
癌症研究
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高影响因子
危险品化学品经营许可证（带存储）