ANALYST
·
2019
·Vol. 144
·期 12
·3817-3825
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IF 4.019
Cuiling Zhang,Dan Li,Dongwei Li,Kai Wen,Xingdong Yang,Ye Zhu
Herein, an ultrasensitive and label-free electrochemical biosensor was developed for microRNA (miRNA) based on rolling circle amplification (RCA)-mediated palladium nanoparticles (PdNPs). The sensor was fabricated by immobilizing dual-functionalized hairpin probes onto an electrode. The specific recognition of target miRNA-21 by the hairpin probes could trigger the RCA reaction, which produced numerous guanine (G)-rich long single-stranded DNAs (ssDNAs). Based on the interaction of the PdII species with the nitrogen atoms of the G bases, these G-rich long ssDNAs served as specific templates in the in situ synthesis of massive PdNPs as electrochemical indicators. The formation of PdNPs was demonstrated to be exactly along the RCA products by high-resolution transmission electron microscopy. Using this cascade signal amplification strategy, the developed biosensor achieved a linear range of 50 aM–100 fM with an ultralow detection limit of 8.6 aM miRNA-21. Furthermore, the developed biosensor exhibited good selectivity, reproducibility, stability and satisfactory feasibility for miRNA-21 detection in human serum samples; this ensured significant potential of this biosensor in disease diagnosis and prognosis applications.
研究领域:
生物传感技术
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纳米医学
·
分子诊断
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DOI
PubMed
高影响因子
JOURNAL OF THE ELECTROCHEMICAL SOCIETY
·
2017
·Vol. 164
·期 9
·B421
·
IF 3.259
Jing Lu,Lin Wu,Yufang Hu,Sui Wang,Zhiyong Guo
A Faraday cage-type electrochemiluminescence (ECL) biosensor for ultrasensitive detection of cancer-associated microRNA-21 was constructed based on Fe3O4@Au magnetic nanoparticles (Fe3O4@Au MNPs) and multi-functionalized graphene oxide. The capture unit (Fe3O4@Au MNPs-cDNA) was prepared by immobilizing capture DNA (cDNA) on Fe3O4@Au MNPs, while the multi-functionalized graphene oxide acted as the signal unit ((sDNA&luminol)@GO). In this biosensor, the capture unit, which was used to catch the microRNA-21, could be easily attracted onto the magnetic glassy carbon electrode surface. After the hybridization of microRNA-21 with the capture unit, the signal unit was further hybridized to form a Faraday cage-type structure, in which the signal unit was directly fixed onto the electrode surface, resembling a huge net and even becoming a part of the electrode surface. All luminol molecules labeled on the signal unit could take part in the electrode reaction, thus greatly enhancing the ECL signal. The ECL intensity was found to increase with the logarithm of microRNA-21 concentrations from 1 fM to 10 pM with a detection limit of 0.3 fM. In addition, this proposed Faraday cage-type ECL biosensor can be used for practical analysis of microRNA-21 in human serum, providing an ultrasensitive detection platform for microRNAs in clinical diagnosis.
研究领域:
生物传感器
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纳米技术
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癌症诊断
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电化学
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分子生物学
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DOI
高影响因子
ACS Applied Materials & Interfaces
·
2015
·
IF 6.723
Cuiyun Yang,Kai Shi,Baoting Dou,Yun Xiang,Yaqin Chai,Ruo Yuan
On the basis of the use of silver nanoclusters (AgNCs) in situ synthesized by cytosine (C)-rich loop DNA templates as signal amplification labels, the development of a label-free and highly sensitive method for electrochemical detection of microRNA (miRNA-199a) is described. The target miRNA-199a hybridizes with the partial dsDNA probes to initiate the target-assisted polymerization nicking reaction (TAPNR) amplification to produce massive intermediate sequences, which can be captured on the sensing electrode by the self-assembled DNA secondary probes. These surface-captured intermediate sequences further trigger the hybridization chain reaction (HCR) amplification to form dsDNA polymers with numerous C-rich loop DNA templates on the electrode surface. DNA-templated synthesis of AgNCs can be realized by subsequent incubation of the dsDNA polymer-modified electrode with AgNO3 and sodium borohydride. With this integrated TAPNR and HCR dual amplification strategy, the amount of in situ synthesized AgNCs is dramatically enhanced, leading to substantially amplified current response for highly sensitive detection of miRNA-199a down to 0.64 fM. In addition, the developed method also shows high selectivity toward the target miRNA-199a. Featured with high sensitivity and label-free capability, the proposed sensing scheme can thus offer new opportunities for achieving sensitive, selective, and simple detection of different types of microRNA targets.
研究领域:
生物化学
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分子生物学
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纳米技术
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分析化学
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DOI
PubMed
高影响因子
ANALYTICA CHIMICA ACTA
·
2016
·
IF 4.712
Yongmei Wu,Kai Sheng,Yongqin Liu,Qian Yu,Baoxian Ye
In this work, an ultrasensitive electrochemical microRNA detection strategy was developed based on porous palladium-modified horseradish peroxidase sphere ( [email protected] ) and target-induced assembly of DNAzyme. A highly loaded HRP sphere was prepared by covalent layer-by-layer assembly with CaCO 3 as sacrificial template for the first time, and was further modified with porous Pd. Notably, [email protected] composite showed a good redox activity of HRP and electrocatalytic activity toward H 2 O 2 . The utilization of [email protected] as electrochemical signal indicator and enhancer to fabricate biosensor could avoid the need for additional redox mediator and amplify the detection sensitivity. Moreover, target recycling amplification was achieved by Pb 2+ -induced cleavage of ternary “Y” structure, circumventing the use of labile nuclease . Subsequent DNA concatamer synthesized through rolling circle amplification (RCA) reaction with cleaved hairpin probe as primer, hybridized with plentiful [email protected] probes, which led to the increased loading of redox-active and electrocatalytic [email protected] for sensitivity improvement. So the proposed electrochemical biosensor detected miRNA-24 down to 0.2 fM (S/N = 3) with a wide linear range from 3 fM to 1 nM. With bifunctional [email protected] tag, DNAzyme-aided target recycle and programmable junction probe, this strategy possessed the advantages of high efficiency, high sensitivity, low cost and versatility, and thus held great promise for other low-abundance nucleic acids determination.
研究领域:
分析化学
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生物化学
·
分子生物学
·
生物医学工程
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PubMed
高影响因子
ACS Applied Materials & Interfaces
·
2018
·
IF 8.097
Ling-Yan Zhai,Min-Xiang Li,Wei-Lun Pan,Yun Chen,Min-Min Li,Jian-Xin Pang,Lei Zheng,Jin-Xiang Chen,Wen-Jun Duan
Breast cancer is the second cause of cancer mortality in women globally. Early detection, treatment, and metastasis monitoring are of great importance to favorable prognosis. Although conventional diagnostic methods, such as breast X-ray mammography and image positioning biopsy, are accurate, they could cause radioactive or invasive damage to patients. Liquid biopsy as a noninvasive method is convenient for repeated sampling in clinical cancer prognostic, metastatic evaluation, and relapse monitoring. MicroRNAs encased in exosomes circulating in biofluids are promising candidate cancer biomarkers because of their cancer-specific expression profiles. Here, we report an in situ detection of microRNA-1246 (miR-1246) in human plasma exosomes as breast cancer biomarker by a nucleic acid functionalized Au nanoflare probe. Needing neither time-consuming and costly isolation of exosomes from the plasma sample nor transfection means, the Au nanoflare probe can directly enter the plasma exosomes to generate fluorescent signal quantitatively by specifically targeting miR-1246. Only 40 μL of plasma is needed to incubate 4 h with the probe, giving signal sensitive enough to distinguish samples of breast cancer to normal control. Using plasma miR-1246 level detected by our assay as a marker, we differentiated 46 breast cancer patients from 28 healthy controls with 100% sensitivity and 92.9% specificity at the best cutoff. This simple, accurate, sensitive, and cost-effective liquid biopsy by the Au nanoflare probe is potent to be developed as a noninvasive breast cancer diagnostic assay for clinical adaption.
研究领域:
肿瘤学
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分子生物学
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纳米技术
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生物医学工程
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DOI
PubMed
高影响因子
ANALYTICAL CHEMISTRY
·
2015
·
IF 5.636
Feifei Xu,Ting Yang,Yun Chen
The distorted and unique expression of microRNAs (miRNAs) in cancer makes them an attractive source of biomarkers. However, one of prerequisites for the application of miRNAs in clinical practice is to accurately profile their expression. Currently available assays normally require pre-enrichment, amplification, and labeling steps, and most of them are semiquantitative. In this study, we converted the signal of target miR-21 into reporter peptide by a DNA-peptide probe and the reporter peptide was ultimately quantified using LC-MS/MS-based targeted proteomics. Specifically, substrate peptide GDKAVLGVDPFR containing reporter peptide AVLGVDPFR and tryptic cleavage site (lysine at position 3) was first designed, followed by the conjugation with DNA sequence that was complementary to miR-21. The newly formed DNA-peptide probe was then hybridized with miR-21, which was biotinylated and attached to streptavidin agarose in advance. After trypsin digestion, the reporter peptide was released and monitored by a targeted proteomics assay. The obtained limit of quantification (LOQ) was 1 pM, and the detection dynamic range spanned ∼5 orders of magnitude. Using this assay, the developed quasi-targeted proteomics approach was applied to determine miR-21 level in breast cells and tissue samples. Finally, qRT-PCR was also performed for a comparison. This report grafted the strategy of targeted proteomics into miRNA quantification.
研究领域:
癌症生物学
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分子生物学
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蛋白质组学
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分析化学
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DOI
PubMed
高影响因子
ANALYTICA CHIMICA ACTA
·
2017
·
IF 4.95
Wenyan Nie,Qing Wang,Xiaohai Yang,Hua Zhang,Zhiping Li,Lei Gao,Yan Zheng,Xiaofeng Liu,Kemin Wang
A high sensitive surface plasmon resonance (SPR) biosensor was proposed based on the gold nanoparticles-decorated molybdenum sulfide (AuNPs-MoS 2 ) for the first time. This SPR platform using the AuNPs-MoS 2 nanocomposites as signal labels for the sensitive and facile measurement of microRNA (miRNA) was executed in only two steps. At first, the thiol-modified DNA oligonucleotide probes , including a sequence complementary to the target miRNA-141, were fixed on the Au film to identify the segment sequence of target miRNA-141. Then, the assistant DNA-linked AuNPs-MoS 2 nanocomposites were used to combine with the other section of the miRNA-141. Performance of SPR biosensor was enhanced by taking advantage of the AuNPs-MoS 2 nanocomposites, thus this newly presented sensing assay exhibited high sensitivity toward miRNA with a detection limit of 0.5 fM. Furthermore, the method showed high specificity, resulting in distinguishing differences among miRNA-200 family members. Especially, the assay could also be used to detect human miRNA from cancer cells, and the results were in excellent accord with the ones obtained using qRT-PCR. What is more, this presented method could be feasible for determining miRNA in 10% human serum. This assay may provide a great potential as a miRNA quantification method in complex samples, and exert significant effect on biomedical research and clinical early diagnosis.
研究领域:
生物医学工程
·
分子生物学
·
纳米技术
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DOI
PubMed
高影响因子
MICROCHIMICA ACTA
·
2019
·Vol. 186
·期 2
·1-9
·
IF 5.479
Jiang Ningjing,Hu Yongjun,Wei Wei,Zhu Tingfeng,Yang Kang,Zhu Guichi,Yu Meng
A surface-enhanced Raman scattering (SERS) method is described for the determination of microRNA that is associated with various forms of cancer. The substrate consists of functionalized gold-silver bimetallic structure, and the sensitivity is strongly enhanced by making use of a re-circulated enzymatic amplification system (REAS). Poly-dopamine acts as both a reductant and a protective of the substrates. It was employed to link the gold core and silver satellite. The unique “hot spots” consisting of a Au@PDA@Ag nanocomposite improve the Raman signal and sensitivity. The reductive feature of PDA can prevent the susceptible oxidation of metallic silver to maintain the high Raman activity. To improve the sensitivity of the assays, a re-circulated enzymatic amplification system was developed in which the nicking endonuclease triggers the nucleic acid reaction system to enter an amplified cycle. By integrating the bimetallic nanosubstrate and magnetic separation into the REAS, microRNA can be detected by SERS (best at the Raman band of 1586 cm −1 ) with a limit of detection as low as 0.2 fM. In our perception, the assay provides an exciting new avenue to study the expression of tumor genes. Thus, it holds vast promise in cancer diagnosis.
Schematic presentation of the SERS method based on poly-dopamine mediated bimetallic SERS substrate and re-circulated enzymatic amplification.
研究领域:
分析化学
·
分子生物学
·
癌症研究
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DOI
PubMed
高影响因子
ANTI-CANCER DRUGS
·
2023
·
IF 2.3
Wang Guangxin,Gao Xia,Sun Zhijun,He Tianyou,Huang Chaogang,Li Shouwei,Long Haocheng
Enhancing research indicatedthat circular RNA (circRNA) acted a critical part in cholangiocarcinoma (CHOL) development. This research aims to discover the role of circRNA SWI/SNF-related, matrix-associated, actin-dependent regulator of chromatin, subfamily a, member 5 ( SMARCA5 ) in CHOL bio-progression, which has been proved to be downregulated in CHOL tissues. In this study, quantitative reverse transcription polymerase chain reaction was used to reveal the level and linkage of circRNA SMARCA5 , miRNA-95-3p and TNF receptor-associated factor 3 gene ( TRAF3 ) in CHOL tissues and cancer cells. The target sites of circRNA SMARCA5 and miRNA-95-3p were forecast by Starbase, and Targetscan was conducted to forecast the potential linkage points of TRAF3 and miRNA-95-3p , and which is affirmed by double luciferase reporter assay. CCK-8 and flow cytometry assay was carried to indicate cell viability. And apoptosis-related protein was counted by caspase3 activity and Western blot assay. CircRNA SMARCA5 was downregulated in CHOL cell lines and cancer samples. Besides, over-expression of SMARCA5 inhibited cell growth and promoted apoptotic rate. Dual-luciferase reporter assays presented that miRNA-95-3p could link with circRNA SMARCA5 . Moreover, miRNA-95-3p was discovered highly expressed in CHOL. Interference of miRNA-95-3p repressed cell proliferation and raised the apoptosis. Importantly, TRAF3 was validated to be a downstream of miRNA-95-3p . Strengthen of miRNA-95-3p reversed the inhibitory impact of circRNA SMARCA5 -plasmid transfection, and the results of miRNA-95-3p inhibitor were reversed by si- TRAF3 . CircRNA SMARCA5 is involved in CHOL development by interosculating miRNA-95-3p / TRAF3 axis and may become a novel approach for treating CHOL.
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PubMed
高影响因子
BIOSENSORS & BIOELECTRONICS
·
2023
·
IF 12.6
Liangliang Wang,Xiufeng Wu,Guanyu Chen,Yawen Chen,Lilan Xu,Jianmin Wang,Jinghua Chen
Exosomal microRNA (miRNA) are important biomarkers for liquid biopsy, and display clinical molecular signatures for cancer diagnosis. Although advanced detection methods have been established to detect exosomal miRNAs, they are faced with certain challenges. Therefore, we aimed to establish a dual amplification-based electrochemical method for detecting exosomal miRNA. This method combined a two-hairpins-based ternary hybridization structure (thTHS)-initiated single-stranded DNA (ssDNA) amplification reaction (ssDAR) and sodium perchlorate (NaClO 4 )-assisted electrocatalytic cycle. Two DNA hairpins were designed to hybridize with target miRNA, forming thTHS. Next, ssDAR was triggered by thTHS to produce long ssDNA on magnetic beads. The long ssDNA, complementary to the signal probes , was subsequently released onto a methylene blue (MB)-labeled double-stranded DNA-modified electrode for strand displacement reaction. This led to a quantitative change in MB and a change in electrocatalytic reduction current from the electrocatalytic cycle of MB-ferricyanide. An amplified electrocatalytic reduction current was produced by adding NaClO 4 to the electrocatalytic system , which substantially improved the signal response range and detection sensitivity. Ultimately, exosomal miRNA detection was achieved by recording changes in the electrocatalytic reduction current before and after miRNA addition. This electrochemical method exhibited a sensitive concentration response with a detection limit of 45 aM and selective miRNA recognition, and successfully used to detect exosomal miRNA derived from cells and serum. Additionally, this method exhibited better discrimination ability between patients with breast cancer (BC) and those people without BC (patients with benign breast disease and healthy people), providing a promising strategy for detecting and monitoring cancer biomarkers.
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高影响因子
BIOSENSORS & BIOELECTRONICS
·
2023
·
IF 12.6
Mingxiang Li,Zhanxiang Liu,Yuchuan Liu,Hu Luo,Ke-Jing Huang,Xuecai Tan
To achieve sensitive detection of low-content microRNA , photoelectrochemical/electrochromic dual-mode sensor with intrinsically low background signal has been developed, but the two detection modules are usually designed with a series-connected structure, which may cause signal interference and thus affect the detection reliability. To solve the above problems, a decoupled dual-mode bioassay for sensitive miRNA-21 detection with high reliability is constructed in this work, by selecting two capacitors to realize parallel amplification for the two detection modules, supplemented with a 3D DNA nanoring photoelectrode signal amplification strategy. The complete decoupling of the two detection modes, photoelectrochemical and electrochromic, as well as the use of digital multimeter , improves the reliability and accuracy of the sensor, and also frees it from dependence on electrochemical workstation, making detection more intuitive and faster. With simple structure, low cost, good reproducibility, high sensitivity, and easy operation, the capacitor-parallel-amplified decoupled photoelectrochemical/electrochromic dual-mode bioassay has broad application prospect in on-site point-of-care detection of diseases and low-cost clinical diagnosis. The design idea of decoupled dual-mode detector can also be extended to the construction of other dual-mode methods.
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高影响因子
ANALYTICA CHIMICA ACTA
·
2022
·
IF 6.911
Yang-Yang Hou,Wan-Zhen Xie,Ke-Jing Huang,Jing Xu
A novel self-powered biosensor is engineered by the integration of DNAzyme walker and AuNPs/graphdiyne biosensing interface, realizing sensitive detection of target microRNA . The cleverly constructed DNAzyme walker with outstanding signal transduction ability to obtain an amplified signal response. In addition, the AuNPs/graphdiyne significantly improves electron transport speed of biosensing interface for improving the sensitivity of biosensor. A dynamic linear range of 0.05 fM-10 pM with a low detection limit of 0.015 fM (S/N = 3) is obtained by utilizing the self-powered biosensor. Meanwhile, the developed self-powered biosensor is capable of assaying miRNA-21 in human serum samples with satisfactory recoveries. This strategy provides a valid method for the sensitive microRNA detection, and shows great potential in point-care detection of tumor biomarker.
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高影响因子
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