Biosensors-Basel
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2023
·Vol. 13
·期 9
·854
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IF 5.4
Yang Qing,Haobin Fang,Yuxing Yang,Yazhen Liao,Haiyu Li,Zhencui Wang,Jie Du
At present, a large number of studies have demonstrated that miRNAs can be used as biological indicators for the diagnosis and treatment of diseases such as tumours and cancer, so it is important to develop a new miRNA detection platform. In this work, miRNA-122 is used as the basis for targeting detection agents. We have designed an unlabelled DNA1 that undergoes partial hybridisation and has a 20 T base long strand. The fluorescent signal in this experiment is derived from copper nanoclusters (CuNCs) generated on the circular T-long strand of DNA1. At the same time, DNA1 is able to react with miRNA-122 and achieve hydrolysis of the part bound to miRNA-122 via the action of nucleic acid exonuclease III (Exo III), leaving a part of the DNA, called DNA3, while releasing miRNA-122 to participate in the next reaction, thus achieving circular amplification. DNA3 is able to react with DNA2, which is bound to streptavidin magnetic beads (SIBs) and separated from the reaction solution via the application of a magnetic field. Overall, this is a fluorescence signal reduction experiment, and the strength of the fluorescence signal from the copper nanoclusters can determine whether the target miRNA-122 is present or not. The degree of fluorescence reduction indicates how much DNA1, and thus the amount of target miRNA-122, has been hydrolysed. By evaluating the variations in the fluorescence signal under optimised conditions, we discovered that this method has good sensitivity, with a detection limit as low as 0.46 nM, better than many other previous works on fluorescence signal-based biosensors for miRNA detection. This technique offers high discrimination and selectivity and can serve as a persuasive reference for early diagnosis.Keywords:exonuclease III;miRNA-122;CuNCs;fluorescence reduction
研究领域:
分子生物学
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生物医学工程
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产品关联
DOI
PubMed
高影响因子
ANALYTICA CHIMICA ACTA
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2021
·
IF 6.558
Kai Tang,Wei Wang,Zhiling Song,Xiliang Luo
A rationally designed multifunctional polydopamine (PDA)-coated metal-organic frameworks (MOFs) biosensors for detection of miRNA-122 with Zn 2+ -triggered aggregation-induced enhancement (AIE) and synergistic chem-photothermal therapy in vitro was developed for the first time. Further, it was successfully used for enhanced fluorescence imaging of miRNA-122 in living cells. The pH-responsive MOFs structure was decomposed under the influence of acidic environment, and a large amount of free Zn 2+ was released as the trigger agent for AIE signal amplification, realizing the ultra-sensitive detection of miRNA-122 and the accurate discrimination of the cells with different expression levels of miRNA-122, with the detection limit as low as 12.5 pM. Meanwhile, ZIF-8 nanoparticles with high loading rate can effectively deliver therapeutic drugs to achieve responsive release. In addition, the modification of versatile PDA-coating provides the biosensor with a faster drug release capability and photothermal conversion performance, demonstrating its superior synergistic chem-photothermal therapy performance. It is expected to play an important role in the integration of cancer diagnosis and synergistic therapy.
研究领域:
生物医学工程
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分子生物学
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肿瘤学
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DOI
PubMed
高影响因子
CHEMOMETRICS AND INTELLIGENT LABORATORY SYSTEMS
·
2022
·
IF 4.175
Qing Liu,Xin Fang,Zeng-Ping Chen,Cai-Xia Shi,Ru-Qin Yu
Herein, the hydrolysis patterns of DNase I against a DNA probe in both single-strand form and DNA/miRNA heteroduplex were experimentally investigated in detail by mass spectrometric technique. Based on the knowledge obtained in the experiments, DNase I-assisted signal amplification strategy was proposed and used to design a mass spectrometric platform for miRNA detection. As a proof of concept, the performance of the proposed platform was evaluated for the detection of miRNA-122 in cell lysate samples. An advanced calibration model was explicitly derived to model the relationships between the concentration of miRNA-122 and the mass spectral signals of mass spectrometrically detected fragments of the DNA probe produced in the DNase I-assisted signal amplification process. With the aid of the advanced calibration model, the proposed mass spectrometric platform achieved quite accurate quantification results for miRNA-122 in cell lysate samples of Huh-7 and HepG-2 cell lines with recovery rates ranging from 98% to 104%. It is noteworthy that the proposed mass spectrometric platform can differentiate the target miRNA from other non-target ones based on their mass spectral response patterns instead of mass spectral intensities, and hence possesses high specificity for the target miRNA. It is reasonable to expect that the combination of mass spectrometry based on DNase I-assisted signal amplification with the advanced chemometric model provides a competitive alternative for designing cost-effective and label-free miRNA detection methods.
研究领域:
生物化学
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分子生物学
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分析化学
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DOI
高影响因子
TALANTA
·
2021
·
IF 6.057
Min-Xi Li,Yao Chen,Zeng-Ping Chen,Ru-Qin Yu
Herein, a label free and sensitive miRNA detection method with enhanced practical applicability was developed based on the locked nucleic acid (LNA) assisted repeated fishing amplification strategy. The working mechanism of the proposed method is as follows: 1) a DNA probe (i.e, L-DNA) with LNA bases is immobilized onto the surface of a gold foil. The L-DNA hybridizes with the 3′ terminus of the first strands of complementary deoxyribonucleic acid (cDNA) of the target miRNA in the test samples; 2) The protruding 5′ terminus of the cDNA serves as a ‘fishhook’ to repeatedly fish the products of a hybridization chain reaction (HCR) out from a ‘reaction tube’; 3) The HCR products can be unloaded from the gold foil into a ‘product tube’ through temperature-controlled dehybridization; 4) The concentration of the target miRNA is determined based on the fluorescence intensity generated by the addition of SYBR-Green I (SG) into the ‘product tube’. The proposed platform was applied to the detection of miRNA-122 in cell lysate samples and obtained quantitative results with accuracy comparable to the quantitative reverse transcription PCR method (qRT-PCR). It is worth pointing out that the proposed platform achieved a limit of detection value of 2.9 fM for miRNA-122 by a simple but effective LNA-assisted repeated fishing amplification strategy instead of complicated enzyme-based amplification techniques. It is reasonable to expect that the proposed method provides a competitive alternative for designing practically applicable, cost-effective and label-free miRNA detection methods.
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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产品关联
DOI
PubMed
高影响因子
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