TALANTA
·
2019
·
IF 4.916
Xiangyu Ma,Haoran Xu,Kun Qian,Martha Kandawa-Schulz,Weimin Miao,Yihong Wang
MicroRNAs (miRNAs) are important biomarkers in early diagnosis of disease. In this work, we developed a simple and effective electrochemical biosensor based on Au nanoparticles (AuNPs)/Carbon nitride nanosheet (CNNS) nanocomposite for the miRNAs detection. Duplex-specific nuclease (DSN) and hairpin structure probe were utilized to improve the sensitivity and selectivity respectively. In the presence of miRNA-21, the signal molecule could be released from the surface of the electrode and decrease the current peak in square wave voltammetry (SWV) test. Under the optimal conditions, the reported biosensor showed that the detection range of miRNA-21 is from 10 fM to 1 nM and detection limit is as low as 2.9 fM. Furthermore, the detection of miRNA-21 added in serum samples indicates the developed biosensor with good selectivity, stability and reproducibility which verify its potential to be used in the early diagnosis of diseases.
研究领域:
生物化学
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分子生物学
·
医学诊断
✓ 已验证
DOI
PubMed
高影响因子
Journal of Materials Chemistry B
·
2018
·Vol. 6
·期 36
·5795-5801
·
IF 4.776
Xuemei Yin,Linlin Liang,Peini Zhao,Feifei Lan,Lina Zhang,Shenguang Ge,Jinghua Yu
MicroRNAs (miRNAs) are a class of significant biomarkers; however, it is still a huge challenge to express them accurately. Herein, a fluorescent/colorimetric dual-model biosensor based upon the quenching effect of graphitic carbon nitride on palladium nanoclusters (Pd NCs) on the platform of a microfluidic paper-based analytical device was built for the detection of miRNAs. On the one hand, Pd NCs could catalyze a chromogenic reaction so that preliminary detection was achieved by the naked eye. On the other hand, the fluorescence analysis combined with nucleic acid cycle signal amplification was required to get precise result and the detection limit is 3 fM, which was superior to the previous method. What's more, this biosensor could be designed to detect other miRNAs via changing the corresponding aptamer sequences. Therefore, the as-constructed biosensor supplies a versatile platform to conduct point-of-care detection of miRNAs with outstanding performance.
研究领域:
生物化学
·
分子诊断
·
纳米技术
·
医学检测
✓ 已验证
DOI
PubMed
高影响因子
ANALYTICAL CHEMISTRY
·
2022
·
IF 8.008
Zizhong Yang,Birong Liu,Ting Huang,Bao-Ping Xie,Wen-Jun Duan,Min-Min Li,Jin-Xiang Chen,Jun Chen,Zong Dai
Sensitive and specific imaging of microRNA (miRNA) in living cells is of great value for disease diagnosis and monitoring. Hybridization chain reaction (HCR) and DNAzyme-based methods have been considered as powerful tools for miRNA detection, with low efficient intracellular delivery and limited amplification efficiency. Herein, we propose a Hairpins@MnO2 nanosystem for intracellular enzyme-free exponential amplification for miRNA imaging. The enzyme-free exponential amplification is based on the synergistic cross-activation between HCR and DNAzymes. The MnO2 nanosheets were employed as the carrier of three kinds of hairpin DNA probes and further provided appropriate Mn2+ as DNAzyme cofactors in the living cell. Upon entering cells and in the presence of highly expressed glutathione (GSH) in tumors, MnO2 is reduced to release Mn2+ and the three kinds of hairpin DNA probes. In the presence of target miRNA, the released hairpin DNA H1 and H2 probes self-assemble via HCR into the wire-shaped active Mn2+-based DNAzymes which further catalyze the cleavage of H3 to generate numerous new triggers to reversely stimulate HCR amplifiers, thus offering tremendously amplified Förster resonance energy transfer readout. The method has a detection limit of 33 fM, which is 2.4 × 104 times lower than that of the traditional HCR system. The developed method also has a high specificity; even miRNAs with a single base difference can be distinguished. Live cell imaging experiments confirmed that this Hairpins@MnO2 nanosystem allows accurate differentiation of miRNA expression of cancer cells and normal cells. The method holds great potential in biological research of nucleic acids.
研究领域:
生物化学
·
分子生物学
·
纳米技术
·
癌症诊断
✓ 已验证
DOI
PubMed
高影响因子
CHEMBIOCHEM
·
2020
·Vol. 21
·期 18
·2667-2675
·
IF 2.576
Mengjia Xu,Pan Fu,Shu Xing,Yang Zhao,Chao Zhao
Graphical Point-of-care cancer catching ? A novel PNA-DNA 2 triplex consisting of a homopurine PNA strand and two homopyrimidine DNA strands is reported for the first time. On the basis of this phenomenon, a simple, inexpensive colorimetric PNA-DNA 2 triple-helix molecular switch-based sensing platform for sensitive and selective detection of miRNAs from cancer cells is demonstrated. Peptide nucleic acids (PNAs), the synthetic DNA mimics that can bind to oligonucleotides to form duplexes, triplexes, and quadruplexes, could be advantageous as probes for nucleic acid sequences owing to their unique physicochemical and biochemical properties. We have found that a homopurine PNA strand could bind to two homopyrimidine DNA strands to form a PNA-DNA 2 triplex. Moreover, the cyanine dye DiSC 2 (5) could bind with high affinity to this triplex and cause a noticeable color change. On the basis of this phenomenon, we have designed a label-free colorimetric sensing platform for miRNAs from cancer cells by using a PNA-DNA 2 triple-helix molecular switch (THMS) and DiSC 2 (5). This sensing platform can detect miRNA-21 specifically with a detection limit of 0.18 nM, which is comparable to that of the THMS-mediated fluorescence sensing platform. Moreover, this colorimetric platform does not involve any chemical modification or enzymatic signal amplification, which boosts its applicability and availability at the point of care in resource-limited settings. The universality of this approach can be simply achieved by altering the sequences of the probe DNA for specific targets.
研究领域:
分子生物学
·
医学诊断
·
分析化学
✓ 已验证
DOI
高影响因子
MICROCHIMICA ACTA
·
2020
·Vol. 187
·期 2
·1-9
·
IF 6.232
Xiang Ling,Zhang Feng,Feng Jian,Chen Chunyan,Cai Changqun
A method is described for the simultaneous determination of hepatocellular carcinoma-associated microRNA-122 and microRNA-199a/b-3p. This probe consists of two kinds of nanomaterials. The first comprises CdTe@CdS core-shell quantum dots which, on excitation at 375 nm give two emissions, with peak wavelengths at 543 (g-QDs) and at 627 nm (r-QDs). The second comprises gold nanoparticles acting as a quencher. In the absence of the target, g-QD-N1 and r-QD-N2 are stable due to the fluorescence stability. With the addition of microRNA-122 and microRNA-199a/b-3p, g-QD-N1 and r-QD-N2 are conjugated to the surface of AuNP-S1/S2 through base complementary pairing. As a result, fluorescence resonance energy transfer (FRET) occurs, resulting in a decrease at 550 nm and 635 nm respectively, which can realize the simultaneous detection of two different microRNAs. Detection is achieved within 50 min. The detection limits (3σ/k) are 0.2 nM for microRNA-122 and 0.5 nM for microRNA-199a/b-3p. The clinical applicability of the assay was demonstrated by detecting microRNAs in human serum and different cell lysates.
Schematic for the simultaneous determination of microRNA-122 and microRNA-199a/b-3p by FRET.
研究领域:
生物化学
·
纳米技术
·
癌症研究
·
分子生物学
✓ 已验证
产品关联
DOI
PubMed
高影响因子
ANALYTICA CHIMICA ACTA
·
2025
·
IF 6
Noshin Afshan,Tao Cheng,Jiabao Yu,Ke Jiao,Lie Li,Jianwei Jiao,Jin Jiao
Background The early breast cancer diagnosis is a great challenge for the treatment success. Traditional FRET biosensors often rely on a one-to-one output pattern, meaning that each target biomarker generates a single fluorescence signal, limiting both the analytical efficiency and diagnostic value. Furthermore, the use of single biomarker for early diagnosis has substantial drawbacks, as it often leads to incomplete or misleading results. The simultaneous detection of miR-155 and miR-105 offers a more robust and clinically relevant diagnostic tool, reducing the likelihood of false positives and allowing for a more accurate assessment of breast cancer at an early stage. Results In this work, a hand in hand catalytic hairpin assembly (CHA) centered FÖrster resonance energy transfer (HCA-FRET) biosensor is designed, which allows two-in-one analysis of breast cancer related two biomarkers. In this system, miR-155 and miR-105, which are involved in regulating the malignant proliferation of breast cancer, are designed as two inputs as the AND DNA logic gate. Hand in hand CHA amplification circuit is effectively triggered under dual miRNA (miR-155 and miR-105), and the resulting significantly high FRET detection signal is measured which satisfies the AND logic gate function for the simultaneous detection of dual miRNA allied with breast cancer. HCA-FRET biosensor is successfully applied to detect miR-155 (0.01–1000 nM) and miR-105 (0.01–1000 nM) with limit of detections as 0.02 and 0.05 pM, respectively. HCA-FRET has excellent selectivity for recognizing single base mismatches and interfering miRNAs, and shows good stability in different matrices. The FRET detection signal recorded from the serum samples of breast cancer patients is found much greater than that of the healthy controls, which effectively distinguishes breast cancer patients from the healthy persons. Significance The study reports a simple, unique HCA-FRET biosensor which undergo dual target miR-155 and miR-105 biomarkers detection enabling early breast cancer diagnosis. Compared with the traditional single marker detection or two signal output analysis methods, this strategy allows more accurate diagnosis of breast cancer patients from normal people. More importantly, through testing the clinical serum samples, we found that this method is expected to be used to distinguish breast cancer patients from normal people. Therefore, the proposed method has great potential applications for nucleic acid based clinical diagnostics.
研究领域:
分子生物学
·
医学诊断
·
生物化学
✓ 已验证
产品关联
DOI
PubMed
高影响因子
SENSORS AND ACTUATORS B-CHEMICAL
·
2020
·
IF 7.1
Xiaoming Ma,Shan He,Huifang Zhang,Xun Li,Jun Xue,Xiaolin Fan,Yusheng Lu,Yiting Chen,Yanjie Zhang,Jianguo Xu
Herein, we have designed a unique oligonucleotide probe named as bumped switchable molecular probe (BS-MB) and employ it to ultrasensitive detection of miRNA-21. The uniqueness is that the bumped portion renders the BS-MB great design flexibility and divides its long stem into two short stems so that it can play the roles of recognition unit, reporting unit, and polymerization primer and template simultaneously. The opening of BS-MB requires a delayed reaction, allowing the system a better-suppressed background without miRNA-21. In contrast, the introduction of miRNA-21 delayed the full opening of BS-MB, inducing a concurrent three-stage amplification for consuming lots of BS-MBs in one amplification event. During the amplification, lots of target analogues of extended miRNA-21 (E-miRNA-21) and cleavaged miRNA-21 (C-miRNA-21) that can be reused as miRNA-21 to cause further fluorescence enhancement are repeatedly generated. The system retains the simplicity (one oligonucleotide probe) and simplifies the detection procedure (one-step detection). Its mix-to-signaling and one-to-many amplification behaviors are superior to conventional MB based amplification, exhibiting a wide linear response and a low detection limit of 8.1 fM. High specificity against even one-base mutation, desirable recovery rates for testing miRNA-spiked serum samples, and well-defined accuracy for classifying clinically related samples are also demonstrated.
研究领域:
分子生物学
·
生物化学
·
纳米技术
·
分析化学
·
生物医学工程
✓ 已验证
产品关联
DOI
高影响因子
JOURNAL OF ELECTROANALYTICAL CHEMISTRY
·
2020
·
IF 3.807
Ziheng Hu,Biying Zhao,Ping Miao,Xialing Hou,Feifei Xing,Yingying Chen,Lingyan Feng
Herein, we present one three-way junction (TWJ) DNA based electrochemical biosensor for sensitive microRNAs detection with locked nucleic acid (LNA) as capture probe and enzyme-free redox cycling signal amplification strategy. LNA owns higher binding affinity and thermal stability compared to normal DNA sequence . In the presence of target microRNA (miRNA-21), novel TWJ structure can be specially triggered with the assistant of methylene blue (MB)-tagged DNA signal probe, thus producing an enhanced electrochemical current. Furthermore, a chemical reductant, tris (2-carboxyethyl) phosphine (TCEP), is adopted to lead a relayed electron transport between solution TCEP and TWJ-linked MB tag and amplified whole signal intensity. Based on the selectivity from target-induced LNA/DNA TWJ structure and enzyme-free MB cycling amplification strategy, the designed electrochemical biosensor exhibits sensitive miRNA-21 detection, with a linear range from 100 aM to 100 pM and a low detection limit of 77 aM. In addition, it can successfully discriminate the differences between the miRNA family members, presenting a promising response to the direct determination of target miRNA-21 in real biological fluids. Our work provides an alternative LNA-based electrochemical biosensor way in the area of miRNA diagnostics and clinical analysis without complicated enzyme/nanomaterials-assisted signal amplification.
研究领域:
生物化学
·
分子诊断
·
分析化学
✓ 已验证
产品关联
DOI
高影响因子
ANALYTICA CHIMICA ACTA
·
2012
·
IF 4.555
Sai Bi,Yangyang Cui,Li Li
Considering the great significance of microRNAs (miRNAs) in cancer detection and typing, the development of sensitive, specific, quantitative, and low-cost methods for the assay of expression levels of miRNAs is desirable. We describe a highly efficient amplification platform for ultrasensitive analysis of miRNA (taking let-7a miRNA as a model analyte) based on a dumbbell probe-mediated cascade isothermal amplification (DP-CIA) strategy. The method relies on the circularization of dumbbell probe by binding target miRNA, followed by rolling circle amplification (RCA) reaction and an autonomous DNA machine performed by nicking/polymerization/displacement cycles that continuously produces single-stranded G-quadruplex to assemble with hemin to generate a color signal. In terms of the high sensitivity (as low as 1 zmol), wide dynamic range (covering 9 orders of magnitude), good specificity (even single-base difference) and easy operation (one probe and three enzymes), the proposed label-free assay is successfully applied to direct detection of let-7a miRNA in real sample (total RNA extracted from human lung tissue), demonstrating an attractive alternative for miRNA analysis for gene expression profiling and molecular diagnostics, particularly for early cancer diagnosis.
研究领域:
分子生物学
·
医学诊断
·
生物化学
✓ 已验证
DOI
PubMed
高影响因子
ACS Applied Materials & Interfaces
·
2015
·
IF 6.723
Xuehong Min,Yuan Zhuang,Zhenyu Zhang,Yongmei Jia,Abdul Hakeem,Fuxin Zheng,Yong Cheng,Ben Zhong Tang,Xiaoding Lou,Fan Xia
We demonstrate an ultrasensitive microRNA detection method based on an extremely simple probe with only fluorogens but without quencher groups. It avoids complex and difficult steps to accurately design the relative distance between the fluorogens and quencher groups in the probes. Furthermore, the assay could accomplish various detection limits by tuning the reaction temperature due to the different activity of exonuclease III corresponding to the diverse temperature. Specifically, 1 pM miR-21 can be detected in 40 min at 37 °C, and 10 aM (about 300 molecules in 50 μL) miR-21 could be discriminated in 7 days at 4 °C. The great specificity of the assay guarantees that the real 21 urine samples from the bladder cancer patients are successfully detected by our method.
研究领域:
癌症研究
·
分子生物学
·
分析化学
✓ 已验证
产品关联
DOI
PubMed
高影响因子
MICROCHIMICA ACTA
·
2017
·Vol. 184
·期 5
·1305-1313
·
IF 4.58
Zhou Lili,Wang Jian,Chen Zhongping,Li Junlong,Wang Ting,Zhang Zhang,Xie Guoming
MicroRNAs (miRNAs) play a considerable role in cancer occurrence and development, and have been identified as promising noninvasive biomarkers. The authors describe a voltammetric method for the determination of the cancer biomarker microRNA-21 (miRNA). It is based on a combination of a universal DNA signal transducer and isothermal target recycling amplification. A hairpin capture probe is bound to the target miRNA to form a duplex structure and to create a toehold in the transducer for initiating the target recycling amplification reaction. In contrast to traditional capture probes, a mismatched site is introduced to improve its ability to capture the target. In order to reduce the complex design procedures of the sequence and widen the applicability of this method, a signal transducer is introduced. Under optimal conditions, response to target miRNA is linear in the 0.5 to 2000 pM concentration range, with a 56 fM. detection limit (at an S/N ratio of 3). In order to characterize the process of target recycling and the stepwise modification of the electrode, real-time fluorescence, agarose gel electrophoresis, cyclic voltammetry, electrochemical impedance spectroscopy and chronocoulometry were used. The results indicate that this isothermal target recycling amplification results in an electrochemical biosensing scheme with wide potential for sensing other bioanalytes.
Schematic illustration of the electrochemical biosensing platform for miRNA-21 detection based on isothermal target recycling amplification and DNA signal transducer triggered strategy.
研究领域:
生物化学
·
分析化学
·
生物传感器
·
癌症研究
✓ 已验证
DOI
高影响因子
MICROCHEMICAL JOURNAL
·
2025
·
IF 5.1
Wu Chen,Mengfan Guo,Zeyu Chen,Ying Sun,Yudian Huang,Yushan Chen,Yanyan Ma,Juan Sun,Bin Qiu,Hui Lin
The overexpression of microRNA-21 (miRNA-21) is closely associated with gastric cancer (GC) and may be used as a biomarker to diagnose GC early. Here, a colorimetry and fluorescence dual-mode biosensor for the detection of miRNA-21 was developed from polydopamine-coated magnetic upconversion nanoparticles (Fe 3 O 4 @UCNP@PDA) and aptamer-modified gold nanoparticles (AuNP-apt). The strong adsorption of single-stranded DNA by PDA results in close proximity between the energy donors (UCNPs) and acceptors (AuNPs), resulting in luminescence quenching of the UCNPs. The miRNA-21 can hybridize with the apt-conjugated AuNPs, and the AuNPs move away from the UCNPs, which results in the luminescence recovery of the UCNPs and the catalytic activity of the AuNPs. After H 2 O 2 and tetrapaminophenylethylene were added, the yellow solution turned green and gradually deepened with increasing miRNA-21 concentration. Under optimal conditions, the detection limits of miRNA-21 by fluorescence and colorimetry assays are 8.90 pM and 185 pM, respectively. In addition, the developed approach was utilized to measure miRNA-21 in serum samples, and the recovery rates ranged from 105.4-107.1 % and 98.5–109.3 %, respectively. The above results indicate great promise for the early diagnosis of GC because of its high sensitivity and selectivity.
研究领域:
生物化学
·
分子诊断
·
纳米医学
✓ 已验证
产品关联
DOI
高影响因子
危险品化学品经营许可证（带存储）