ANALYTICAL CHEMISTRY
·
2026
·
IF 7.3
Jiayi Hu,Lin Hao,Meiyu Chen,Yufan Zhang
The efficient and quantitative detection of miRNA-21 plays a critical role in early cancer diagnosis, as well as in the screening and treatment of early-stage infectious diseases. In this study, we developed an electrochemical biosensor for the detection of miRNA-21 in human serum samples. The sensor is constructed based on carboxyl-modified cobalt-doped nitrogen-doped carbon (Co-NC–COOH), a dual-output toehold-mediated strand displacement (TMSD) reaction, and gold nanoparticles (Au NPs) to achieve electrocatalytic signal amplification toward acetaminophen (AP). Co-NC–COOH possesses abundant active and catalytic sites, along with a porous surface morphology, which facilitates electron transfer and enhances the loading capacity of the capture probe (CP). In the presence of miRNA-21, the dual-output TMSD reaction is initiated, resulting in the release of a large amount of product strand (P). Subsequently, P hybridizes with the CP immobilized on the electrode surface, enabling the attachment of Au NPs to the electrode and thereby achieving significant electrochemical signal amplification. The results demonstrate that the proposed biosensor exhibits excellent analytical performance, including a wide linear detection range (0.1 fM–50 nM), an ultralow detection limit (63 aM), and satisfactory spike recovery rates (95.18%–103.59%). Moreover, the biosensor shows good stability and high specificity, providing a reliable platform for the precise quantification of miRNA-21 in human serum.
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DOI
PubMed
高影响因子
ANALYTICAL CHEMISTRY
·
2025
·
IF 6.7
Wenting Cheng,Han Pan,Jinhua Chen,Miao He,Zhongyun Wang,Yang Xiang
Exosomal miRNAs, particularly miRNA-21, are promising cancer biomarkers. Current enzyme-dependent detection methods face challenges, such as environmental limitations and high costs. In contrast, enzyme-independent sensors are highly desirable for on-site, miniaturized, and cost-effective miRNA detection. To address these limitations, we developed a nonenzymatic electrochemical sensor featuring a triple-signal amplification system for ultrasensitive detection of miRNA-21. This sensor utilizes cascade toehold-mediated strand displacement reactions to activate molecular machines triggered by target miRNA, generating biotinylated-and-thiol-modified double-stranded DNA for stable immobilization on a gold electrode. Preprepared biotinylated tetrahedron DNA (TDNA)-mediated hybridization chain reaction probes are then linked to the electrode via streptavidin–biotin binding. This amplification process allows for significant DNA duplex immobilization, with electroactive [Ru(NH3)6]3+ (RuHex) adsorbed onto them, producing a robust electrochemical signal. This approach enables accurate detection of miRNA-21 at concentrations as low as 0.43 fM, with a linear range from 1 fM to 1 nM. Clinical testing demonstrates its potential for cancer diagnostics.
研究领域:
生物化学
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医学诊断
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分子生物学
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PubMed
高影响因子
ANALYTICA CHIMICA ACTA
·
2020
·
IF 5.977
Lili Yang,Hao Wang,Haitao Lü,Ni Hui
A rapid, sensitive and low-fouling sensor for application to detect clinical biomarkers directly in biological media is highly desirable. Herein, we report a versatile strategy to prepare polyaniline (PANI) based conducting polymer hydrogel through the assembly of PANI and phytic acid (PA) by dynamic boronate bond. PANI/PA, a conductive hydrogel with high specific area and multiple pore structures have demonstrated excellent antifouling ability and electrochemical property. The electrochemical biosensors for microRNA can be developed by the immobilization of DNA probes onto PANI/PA interface (microRNA24 is used as a model case), and redox currents of PANI were utilized as the sensing signals to assay DNA/RNA hybridization reaction. Owing to the typical characteristics of PANI/PA hydrogel, the biosensor has shown excellent sensing performances with wide linear range (1.0 fM-1.0 pM), low sensing limit (0.34 fM) and efficient ability to detect microRNA mismatches. Given the facile processability, excellent stability and good antifouling property of the PANI/PA hydrogel, the proposed hydrogel-based biosensor may find broad applications in clinical diagnostics and biomedical devices.
研究领域:
材料科学
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生物化学
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分析化学
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生物医学工程
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高影响因子
Oncotarget
·
2017
·
IF 5.168
Hai-Ning Liu,Hao Wu,Yan-Jie Chen,Yu-Jen Tseng,Enkhnaran Bilegsaikhan,Ling Dong,Xi-Zhong Shen,Tao-Tao Liu
Background Many new diagnostic biomarkers have been developed for hepatocellular carcinoma (HCC). We selected two methods with high diagnostic value, the detection of serum microRNAs and metabolomics based on gas chromatography/mass spectrometry (GC/MS), and attempted to establish appropriate models. Methods We reviewed the diagnostic efficiencies of all microRNAs identified by previous diagnostic tests. Then we chose appropriate microRNAs to validate the diagnostic efficiencies, and determined the optimal combination. We included 66 patients with HCC and 82 healthy controls (HCs) and detected the expression of the microRNAs. GC/MS analysis was performed, and we used three multivariate statistical methods to establish diagnostic models. The concentration of alpha feto-protein (AFP) was determined for comparison with the novel models. Results 82 published studies and 92 microRNAs were ultimately included in this systematic review. Seven microRNAs were selected for further validation of their diagnostic efficiencies. Among which, miR-21, miR-106b, miR-125b, miR-182 and miR-224 had a significantly different expression in HCC patients. The combination of miR-21, miR-106b and miR-224 had the highest area under the curve (AUC) at 0.950 with a sensitivity of 80.3% and a specificity of 92.7%. The GC/MS analysis exhibited an excellent diagnostic value and the AUC reached 1.0. In comparison, the AUC of the traditional biomarker, AFP, was 0.755. Conclusion MicroRNAs and metabolomics shows promising potential as new diagnostic methods due to their high diagnostic value compared with traditional biomarkers.
研究领域:
肿瘤学
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分子生物学
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代谢组学
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PubMed
高影响因子
ANALYTICA CHIMICA ACTA
·
2022
·
IF 6.911
Yunyun Ning,Cuiling Zhang,Chunyan Wang,Chuqing Zhou,Nana Gong,Qiang Wang,Ye Zhu
Accurate detection of microRNA (miRNA) is challenging but essential for disease diagnostics and therapeutics. To meet the high demands of miRNA analysis, we proposed a highly sensitive and specific electrochemical strategy based on duplex-specific nuclease (DSN)-assisted target recycling and DNA self-assembled FeN x C nanocatalytic network for precise quantitation of miRNA-21 as a model. FeN x C nanospherical catalyst with high peroxidase-mimicking activity was synthesized through high-temperature pyrolysis and modified with DNA. The identification of miRNA-21 by elaborately designed capture probe triggered DSN-assisted target recycling. Subsequently, self-assembled FeN x C nanocatalytic network was constructed on the sensor surface through DNA hybridization reaction, whose signal response was demonstrated to be 4.272-fold higher than the single layer of FeN x C. By virtue of such cascade signal amplification, ultrasensitive electrochemical detection of miRNA-21 was realized in a dynamic range of 0.5 fM - 1 pM with a detection limit of 0.2805 fM. Additionally, the proposed strategy exhibited high specificity for miRNA-21 and practical applicability in real sample analysis, demonstrating promising potential for the reliable miRNA analysis in disease diagnosis and therapy applications.
研究领域:
生物化学
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分子诊断
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纳米医学
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分析化学
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PubMed
高影响因子
ACS Nano
·
2022
·
IF 18.027
Wei Wei,Huiting Lu,Wenhao Dai,Xiaonan Zheng,Haifeng Dong
Multiplexed profiling of microRNAs’ subcellular expression and distribution is essential to understand their spatiotemporal function information, but it remains a crucial challenge. Herein, we report an encoding approach that leverages combinational fluorescent dye barcodes, organelle targeting elements, and an independent quantification signal, termed Multiplexed Organelles Portrait Barcodes (MOPB), for high-throughput profiling of miRNAs from organelles. The MOPB barcodes consist of heterochromatic fluorescent dye-loaded shell–core mesoporous silica nanoparticles modified with organelle targeting peptides and molecular beacon detection probes. Using mitochondria and endoplasmic reticulum as models, we encoded four Cy3/AMCA ER-MOPB and four Cy5/AMCA Mito-MOPB by varying the Cy3 and Cy5 intensity for distinguishing eight organelles’ miRNAs. Significantly, the MOPB strategy successfully and accurately profiled eight subcellular organelle miRNAs’ alterations in the drug-induced Ca2+ homeostasis breakdown. The approach should allow more widespread application of subcellular miRNAs and multiplexed subcellular protein biomarkers’ monitoring for drug discovery, cellular metabolism, signaling transduction, and gene expression regulation readout.
研究领域:
细胞生物学
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分子生物学
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纳米技术
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药物发现
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生物医学工程
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DOI
PubMed
高影响因子
ANALYTICAL CHEMISTRY
·
2023
·
IF 7.4
Xi Long,Tong Luo,Panpan Yuan,Yuqing Gan,Hui Liu,Zhiwei Deng,Jiacheng Ding,Zan Gong,Yanjing Yang,Shian Zhong
The ability to simply, selectively, and sensitively detect low numbers of miRNAs in clinical samples is highly valuable but remains a challenge. In this work, we present a novel miRNA detection system by using the elaborately designed hairpin switch, where the T7 primer, template, target recognize sequence, and light-up RNA aptamer template are edited and embedded in one single-stranded DNA hairpin structure. In the beginning, the hairpin switch maintained the hairpin structure 1, in which the ds promoter of T7 polymerase was disrupted, thus the transcription reaction of T7 polymerase was inhibited. After binding to the target, the hairpin switch 1 was unfolded and turned to the hairpin structure 2. This switch initiates the in vitro T7 transcription reaction, producing plenty of RNA transcripts containing RNA aptamers. Consequently, transcribed tremendous RNA aptamers lighted up the fluorophore for quantitative analysis. Compared with the existing T7 polymerase-based amplification system, this strategy exhibits several advantages, including simplicity, convenience, and high selectivity and sensitivity. The experimental results demonstrated that we could achieve the quantification of miRNA in buffer and complex biological samples.
研究领域:
分子生物学
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生物技术
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纳米技术
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临床诊断
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PubMed
高影响因子
PEDIATRIC RESEARCH
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2023
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IF 3.6
Wang Qi,Shi Xulai,Li Ping-Ping,Gao Li,Zhou Yueyuan,Li Luyao,Ye Hao,Fu Xiaoqin,Li Peijun
Background Although previous studies show that microRNAs (miRNAs) can potentially be used as diagnostic markers for epilepsy, there are very few analyses of pediatric epilepsy patients. Methods miRNA profiles using miRNA-seq was performed on plasma samples from 14 pediatric epileptic patients and 14 healthy children. miRNA miR-27a-3p that were significantly changed between two groups were further evaluated. The potential target genes of miR-27a-3p were screened through unbiased mRNA-seq and further validated using Western blot and immunohistochemistry in HEK-293T cells and in the brains of mice with epilepsy induced by lithium chloride–pilocarpine. Results We found 82 upregulated and 76 downregulated miRNAs in the plasma from pediatric patients compared with controls ( p < 0.01), of which miR-27a-3p exhibited a very low p value ( p < 0.0001) and validated in additional plasma samples. Two genes, GOLM1 and LIMK1, whose mRNA levels were decreased ( p < 0.001) with the increase of miR-27a-3p were further validated in both HEK-293T cells and in epileptic mice. Conclusions MiR-27a-3p exhibits potential as a diagnostic and therapeutic marker for epilepsy. We postulate that additional studies on the downstream targets of miR-27a-3p will unravel its roles in epileptogenesis or disease progression. Impact
A total of 158 differentially expressed miRNAs were detected in plasma between epileptic and control children.
Plasma miR-27a-3p was one of the miRNAs with a low p value.
GOLM1 and LIMK1 were validated as downstream target genes of miR-27a-3p.
miR-27a-3p has potential as a diagnostic and therapeutic marker for epilepsy.
研究领域:
神经科学
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分子生物学
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儿科
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生物标志物研究
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高影响因子
Journal of Materials Chemistry B
·
2017
·Vol. 5
·期 34
·7133-7139
·
IF 4.543
Yike Fu,Tong Chen,Gang Wang,Tongxu Gu,Congkun Xie,Jie Huang,Xiang Li,Serena Best,Gaorong Han
MicroRNAs (miRNAs) play a key role in regulating gene expression but can be associated with abnormalities linked to carcinogenesis and tumor progression. Hence there is increasing interest in developing methods to detect these non-coding RNA molecules in the human circulation system. Here, a novel FRET miRNA-195 targeting biosensor, based on silica nanofibers incorporated with rare earth-doped calcium fluoride particles (CaF2:Yb,Ho@SiO2) and gold nanoparticles (AuNPs), is reported. The formation of a sandwich structure, as a result of co-hybridization of the target miRNA which is captured by oligonucleotides conjugated at the surface of CaF2:Yb,Ho@SiO2 fibers and AuNPs, brings the nanofibers and AuNPs in close proximity and triggers the FRET effect. The intensity ratio of green to red emission, I541/I650, was found to decrease linearly upon increasing the concentration of the target miRNA and this can be utilized as a standard curve for quantitative determination of miRNA concentration. This assay offers a simple and convenient method for miRNA quantification, with the potential for rapid and early clinical diagnosis of diseases such as breast cancer.
研究领域:
生物传感器
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纳米技术
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分子生物学
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癌症研究
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分析化学
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DOI
PubMed
高影响因子
Frontiers in Pharmacology
·
2023
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IF 5.6
Zhang Xinxin,Xu Min,Cai Shuilin,Chen Bei,Lin Hetong,Liu Zhiyu
Introduction: The protective effects of astaxanthin against myocardial ischemia-reperfusion injuries are well documented, although the mechanisms are not defined.Methods: The anoxia-reoxygenation injury model was established after astaxanthin treated H9c2 cells for 24 h. Cell viability, lactate dehydrogenase, oxidative stress level and western blot were tested. Secondly, measured the effects of astaxanthin pretreatment on microRNA expression in a rat myocardial cell anoxia-reoxygenation injury model.Results: After anoxia-reoxygenation injury, in a dose dependent manner, astaxanthin increased cell viability, superoxide dismutase and glutathione peroxidase activity, decreased lactate dehydrogenase and malondialdehyde levels, downregulated protein expression of caspase-3, caspase-8, nuclear factor erythroid-2-related factor 2 and heme oxygenase-1, and upregulated the Bcl-2/Bax ratio. High-throughput sequencing and qPCR showed that microRNAs rno-miR-125b-5p and rno-let-7c-1-3p were differentially expressed (|log2| ≥ 0.585, q < 0.1) between the normal, anoxia-reoxygenation, and astaxanthin (1.25 μM) groups. Kyoto Encyclopedia of Genes and Genomes and GO Gene ontology pathway enrichment analyses showed that TNF signaling, axon guidance, NF-κB signaling pathway, and other pathways displayed differentially expressed microRNA target genes associated with myocardial injuries.Discussion: These results suggested that thetarget genes of rno-miR-125b-5p were enriched in inflammation and apoptosis-related signaling pathways. Also, the results imply that simultaneous targeting of these related signaling pathways could significantly prevent myocardial anoxia-reoxygenation injury in the presence of astaxanthin.
研究领域:
心血管生物学
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分子生物学
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药理学
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DOI
PubMed
高影响因子
MICROCHEMICAL JOURNAL
·
2026
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IF 5.2
Feifei Zhou,Yingzhan Fan,Qinrui Cao,Yingying Yuan,Yingxiao Chen,Lingling Liu,Jingwen Mai,Nandi Chen,Tianxing Ji,Dou Wang,Chenchen Ge
Dual-function DNA probe (dfDNA) enables a split crRNA/Cas12a (SCas12a) assay. • The dfDNA acts as both a Cas12a activator and a fluorescent reporter. • The dfDNA activates both the cis - and trans -cleavage activities of Cas12a. • The dfDNA/SCas12a assay detects microRNA-21 (miR-21) without amplification. • Bead-assisted dfDNA format improves the sensitivity for miR-21 detection. MicroRNA-21 (miR-21) is a promising non-invasive biomarker for tumor diagnosis, but its short length and low abundance make amplification-free detection challenging. Here, we developed a dual-function DNA probe (dfDNA)-enabled split crRNA-Cas12a (SCas12a) system for miR-21 detection. This design separates crRNA into a constant scaffold fragment (sc-crRNA) and a variable spacer fragment (sp-crRNA), and miR-21 functionally replaces the spacer fragment to activate Cas12a in the presence of dfDNA. The dfDNA acts as both a target-recognition probe and a fluorescent reporter: it hybridizes with miR-21, activates Cas12a, and presents a single-stranded fluorescent overhang that can be cleaved after Cas12a activation. Because target recognition and signal reporting are integrated within a single probe, dfDNA positions the activated Cas12a complex close to the cleavable overhang, improving signal conversion compared with the conventional single-stranded DNA reporter and fully complementary double-stranded DNA probe format. Under optimized conditions, the limit of detection (LOD) with the homogeneous ssDNA reporter-based assay was 347.9 pM for miR-21, whereas the homogeneous dfDNA-based assay reduced the LOD to 19.8 pM. When transferred to a magnetic bead (MB)-assisted format, the dfDNA-based system achieved an LOD of 1.1 pM, supporting that the localized reaction accelerates the trans -cleavage kinetics of Cas12a. The MB-assisted dfDNA assay achieves comparable or even superior sensitivity for miR-21 detection compared with previously reported split crRNA-based fluorescence methods. Further optimization, including target enrichment or signal amplification strategies, is needed before this method can be reliably used for low-abundance miRNAs in clinical serum samples. Download: Download high-res image (88KB) Download: Download full-size image
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高影响因子
BIOSENSORS & BIOELECTRONICS
·
2026
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IF 11.8
Jingtao Huang,Yu Ting Xu,Jin Chen Liu,Wen Yi Lv,Li Ping Cao,Hong Yan Zou,Chun Mei Li,Cheng Zhi Huang
The 10-23 DNAzyme is a potent catalytic nucleic acid for biosensing; however, traditional single-blocker designs suffer from high background signal leakage due to spontaneous thermal “breathing.” To overcome this, we engineered a triangular DNA probe-DNAzyme (TDPzyme) that spatially separates the enzyme components onto a rigid 3D scaffold. By splitting the binding arms and catalytic center, the TDPzyme functions as a fault-tolerant logic “AND” gate, strictly requiring the simultaneous displacement of multiple blocker strands by the target miR-21. This architectural constraint physically shields the probe from nuclease degradation and virtually eliminates false-positive activation, yielding a near-zero background state. Functionalized with a Cy3-BHQ2 pair and an AS1411 aptamer for targeted, carrier-free cellular delivery, the TDPzyme achieved an approximately 18-fold enhancement in sensitivity over conventional closed-arm designs, establishing a linear range of 0.1 nM to 40.0 nM and a limit of detection of 50.3 pM (3σ). Furthermore, time-lapse dynamic imaging demonstrated that the TDPzyme enables highly reliable, high-contrast discrimination of miR-21 expression between tumor and normal cells. Ultimately, the TDPzyme design offers a robust, structurally controllable platform for developing highly sensitive, low-background biosensors applicable to demanding live-cell imaging and precision molecular diagnostics.
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高影响因子
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