Nano Research
·
2024
·
IF 9.5
Sun Yudie,Han Yunxiang,Wang Mingyue,Ye Mingfu,Wu Konglin,Zhang Kui
Circulating microRNAs (miRNAs) play a pivotal role in the occurrence and development of acute myocardial infarction (AMI), and precise detection of them holds significant clinical implications. The development of luminol-based luminophores in the field of electrochemiluminescence (ECL) for miRNA detection has been significant, while their effectiveness is hindered by the instability of co-reactant hydrogen peroxide (H 2 O 2 ). In this work, an iron single-atom catalyst (Fe-PNC) was employed for catalyzing the luminol-O 2 ECL system to achieve ultra-sensitive detection of myocardial miRNA. Target miRNA triggers a hybridization chain reaction (HCR), resulting in the generation of a DNA product featuring multiple sticky ends that facilitate the attachment of Fe-PNC probes to the electrode surface. The Fe-PNC catalyst exhibits high promise and efficiency for the oxygen reduction reaction (ORR) in electrochemical energy conversion systems. The resulting ECL biosensor allowed ultrasensitive detection of myocardial miRNA with a low detection limit of 0.42 fM and a wide linear range from 1 fM to 1.0 nM. Additionally, it demonstrates exceptional performance when evaluated using serum samples collected from patients with AMI. This work expands the application of single-atom catalysis in ECL sensing and introduces novel perspectives for utilizing ECL in disease diagnosis.
研究领域:
生物传感器
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电化学
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纳米技术
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心脏病学
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分子诊断
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DOI
高影响因子
MICROCHIMICA ACTA
·
2024
·Vol. 191
·期 7
·1-7
·
IF 5.3
Wang Shirong,Wei Zaiwa,Li Liangxian,Luo Yu,Huang Zhimei,Yang Xing,Tang Yafang
CRISPR-Cas12a with robust trans-cleavage activity were employed to mitigate background fluorescence signal, achieving sensitive detection of miRNA-21. The activation of trans-cleavage activity of Cas12a was achieved by utilizing cDNA as a trigger. Upon the presence of target miRNA-21, cDNA hybridizes with it forming a DNA/RNA double-stranded structure. Exonuclease III (ExoIII) facilitates the degradation of cDNA, releasing the target for subsequent cycles. Due to cDNA degradation, the trans-cleavage activity of Cas12a remains unactivated and does not disrupt the synthesis template of copper nanoparticles. Addition of Cu 2+ and AA leads to the formation of highly fluorescent copper nanoparticles. Conversely, in absence of miRNA-21, intact cDNA activates trans-cleavage activity of Cas12a, resulting in degradation of the synthesis template and failure in synthesizing fluorescent copper nanoparticles. This method exhibits excellent selectivity with a low limit of detection (LOD) at 5 pM. Furthermore, we successfully applied this approach to determine miRNA-21 in cell lysates and human serum samples, providing a new approach for sensitive determination of biomarkers in biochemical research and disease diagnosis. Graphical Abstract
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高影响因子
RSC Advances
·
2018
·Vol. 8
·期 13
·6781-6788
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IF 2.936
Aixian Zheng,Xiaolong Zhang,Yanbing Huang,Zhixiong Cai,Xiaolong Liu,Jingfeng Liu
MicroRNAs play an important role in various biological processes, and their aberrant expression is closely associated with various human diseases, especially cancer. Real-time monitoring of microRNAs in living cells may help us to understand their role in cellular processes, which can further provide a basis for diagnosis and treatment. In this study, polydopamine was used to assist the versatile modification of a nucleic acid probe for intracellular microRNA imaging and enhanced photothermal therapy. Polydopamine can be covalently linked with a thiol-terminated nucleic acid probe through the Michael addition reaction under slightly alkaline conditions. This modification is mild and can be performed directly in an aqueous solution, which can better resist hydrolysis than the traditional modification processes, resulting in a nanoprobe with better stability and higher loading of nucleic acids. This prepared nanoprobe can easily enter cells without transfection agents and then realize the imaging of intracellular miRNA through fluorescence restoration. Moreover, the coating of PDA can enhance the photothermal conversion efficiency of the nanoprobe, making it suitable for photothermal therapy of cancer. It is expected that the PDA-based versatile modification can help to construct a promising platform for tumor imaging and treatment.
研究领域:
分子生物学
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癌症治疗
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纳米医学
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PubMed
高影响因子
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
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分子诊断
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DOI
PubMed
高影响因子
JOURNAL OF THE ELECTROCHEMICAL SOCIETY
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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
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2015
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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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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
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分子生物学
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生物医学工程
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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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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
·
分子生物学
·
蛋白质组学
·
分析化学
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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
ANALYTICAL CHEMISTRY
·
2021
·
IF 6.986
Ting Bao,Ruobing Fu,Yuying Jiang,Wei Wen,Xiuhua Zhang,Shengfu Wang
The development of a robust sensing platform with an efficient probe assembly, and ingenious signal conversion is of great significance for bioanalytical application. In this work, a multipedal polydopamine nanoparticles–DNA (PDANs-DNA) nanomachine coupling electrochemical-driven metal–organic frameworks (MOFs) conversion-enabled biosensing platform was constructed. The PDANs-DNA nanomachine was designed based on Ca2+-mediated DNA adsorption and target-triggered catalytic hairpin assembly on PDANs, which not only maintained the DNA immobilization simplicity but also possessed a high walking efficiency. PDANs-DNA nanomachine could walk fast on the electrode via multiple legs under exonuclease III driving, resulting in the formation of DNA dendrimers through two hairpins assembly. The MOFs (Fe-MIL-88-NH2) probe was decorated on the DNA dendrimers to act as a porous metal precursor and converted into electroactive Prussian Blue by a controlled electrochemical approach, which was a facile, simple, and room-temperature approach compared with the commonly employed MOFs conversion methods. Using microRNA-21 (miRNA-21) as the model target, the proposed biosensor achieved miRNA-21 detection ranging from 10 aM to 10 pM with the detection limit of 5.8 aM. The proposed strategy presented a highly efficient walking platform with the ingenious electrochemical conversion of MOFs, providing more options for the design of an electrochemical platform and holding potential applications in clinical analysis and disease diagnosis.
研究领域:
生物分析化学
·
纳米生物学
·
医学诊断
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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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高影响因子
危险品化学品经营许可证（带存储）