CHEMICAL RESEARCH IN CHINESE UNIVERSITIES
·
2017
·Vol. 34
·期 1
·28-32
·
IF 1.024
Zhang Hong,Zou Li,Li Ruimin,Zhao Mingqin,Ling Liansheng
Simian virus 40(SV40) is a polyomavirus and can induce a series of different tumors. The recognition of SV40 genome is crucial to tumor diagnosis and gene therapy. Herein, a sensitive and selective colorimetric method for sequence-specific recognition of homopyrimidine·homopurine duplex DNA(dsDNA) of SV40(4424—4440, gp6) was established with a hairpin probe based upon the formation of triplex DNA. Hairpin probe 5′-CCC TAC CCA TTT TTT CTT CTC TTT CCT GGG TAG GGC GGG TTG GG-3′(HP) containing G-rich sequence and 17-bp trip-lex-forming sequence was used as the signal probe, which was stem-loop structure alone and exhibited low catalytic activity. Upon its binding to the target duplex of SV40, hairpin probe transferred from stem-loop structure to parallel triplex DNA, accompanied by the recovery of catalytic activity of DNAzyme and a sharp increase of absorbance. Under optimum conditions, the absorbance was increased proportionally to the concentration of dsDNA over the range from 500 pmol/L to 40.0 nmol/L with a detection limit of 433 pmol/L. Moreover, satisfied results were obtained when the assay was used to recognize the mismatched sequences.
研究领域:
分子生物学
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肿瘤诊断
·
基因治疗
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产品关联
DOI
高影响因子
BIOSENSORS & BIOELECTRONICS
·
2017
·
IF 7.78
Yiyi Tao,Dan Yin,Mingchao Jin,Jie Fang,Tao Dai,Yi Li,Yuxia Li,Qinli Pu,Guoming Xie
An electrochemical microRNA (miRNA) analysis platform by combining double-loop hairpin probe (DHP) and doxorubicin-loaded gold nanoparticles ( [email protected] ) for ultrasensitive miRNA detection is proposed. Firstly, we here report a DHP that is simultaneously engineered to incorporate a miRNA recognition sequence, an output segment and output's complementary fragment. The important aspect of this hairpin probe is that it would not be degraded by duplex specific nuclease (DSN) and circumvents elaborately chemical modification disadvantages encountered by classic molecular beacon . For the DHP-based DSN signal amplification system, DHP hybridizes with target miRNA to form DNA-miRNA heteroduplexes , and the DSN can hydrolyze the DNA in the heteroduplexes structure selectively, while released target miRNA strand can initiate another cycle resulting in a significant signal amplification and the accumulated output segments could be responsible for strand displacement on the electrode directly. Furthermore, a great deal of doxorubicin (Dox) are loaded on the gold nanoparticles (AuNPs) to fabricate the [email protected] biocomposites that could magnify the electrochemical signal and enable the ultrasensitive analysis of miRNA. As a result, the miRNA was capable of being detected in a limit of 0.17 pM and other kinds of miRNA were discriminated facilely by this method. The described DHP as a toolbox and the nano-biocomposites as a novel signal material would not only promote the design of electrochemical biosensors but also open a good way to promote the establishment of test method in malignant tumors.
研究领域:
生物化学
·
分子生物学
·
纳米医学
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DOI
PubMed
高影响因子
SENSORS AND ACTUATORS B-CHEMICAL
·
2023
·
IF 8.4
Weihua Zhao,Jun Xu,Hongbo Li,Shengliang Zhong,Ruqin Yu
MiRNA can exist in body fluids and its changes directly affect human life and health, which has been a research hotspot in recent years. In this work, we designed an enzyme-free sensing system for highly sensitive detection of miRNA-21 based on strand displacement reaction (SDR) that can recycle the target and functional hairpin probes (HP) that modified on upconversion nanomaterials (UCNPs). The target gene circulates through the SDR process and completes the initial signal amplification process. Then, the C2 sequences and targets released by the SDR process can continue to be combined with functional hairpin probes modified with fluorophore and quenching groups. After infrared irradiation, HP is broken, and C2 or the target can take the fluorophores away from the quenching group, so that the fluorescence signal can be detected to achieve highly sensitive detection of the target. In sensing system, the use of magnetic beads can reduce the background signal during the reaction process, and the functional hairpin probe can accurately turn the signal on and off. The sensor system not only shows excellent detection performance (detection limit of 30.3 fM) in buffer and actual detection, but also has excellent selectivity, which can provide a new detection method for miRNA-21 detection.
研究领域:
生物化学
·
分子生物学
·
纳米医学
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产品关联
DOI
高影响因子
ANALYTICAL CHEMISTRY
·
2024
·
IF 6.7
Feifei Sun,Jinde Liu,Zhuoqun Su,Di Wu,Shaoqi Qu,Yongning Wu,Lin Li,Guoliang Li
Owing to the co-occurrence of hazardous compounds, it is crucial to build multiple highly discriminative probe libraries for simultaneous determination. Drawing inspiration from nucleic acid barcodes, we developed a probe system that is exclusively based on the nucleic acid secondary structure’s hairpin structure, which can be directly read by nanopores. The highly distinguishable hairpin probes were constructed, and a detailed explanation of the possible patterns in their design was provided. These probe-representative events measured through the α-hemolysin (α-HL) nanopores were both distinguished, either through visual observation or comparison of the nanopore parameters. Besides, the potential design pattern for probes with unique telegraphic switching between the two levels was also unveiled. Finally, these probes were utilized to realize simultaneous, ultrasensitive mycotoxin multiple-detection, and their prospective applications for the detection of proteins and microRNAs were presented, indicating their suitability for a wide range of sensing applications.
研究领域:
生物技术
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分子生物学
·
分析化学
✓ 已验证
产品关联
DOI
PubMed
高影响因子
ANALYTICAL CHEMISTRY
·
2024
·
IF 6.7
Junhua Chen,Xu Wang,Junhui Guo,Yiwen Lv,Manjia Chen,Hui Tong,Chengshuai Liu
The uranyl ion (UO22+) is the most stable form of uranium, which exhibits high toxicity and bioavailability posing a severe risk to human health. The construction of ultrasensitive, reliable, and robust sensing techniques for UO22+ detection in water and soil samples remains a challenge. Herein, a DNA network biosensor was fabricated for UO22+ detection using DNAzyme as the heavy metal recognition element and double-loop hairpin probes as DNA assembly materials. UO22+-activated specific cleavage of the DNAzyme will liberate the triggered DNA fragment, which can be utilized to launch a double-loop hairpin probe assembly among Hab, Hbc, and Hca. Through multiple cyclic cross-hybridization reactions, hexagonal DNA duplex nanostructures (n[Hab•Hbc•Hca]) were formed. This DNA network sensing system generates a high fluorescence response for UO22+ monitoring. The biosensor is ultrasensitive, with a detection limit of 2 pM. This sensing system also displays an excellent selectivity and robustness, enabling the DNA network biosensor to work even in complex water and soil samples with excellent accuracy and reliability. With the advantages of enzyme-free operation, outstanding specificity, and high sensitivity, our proposed DNA network biosensor provides a reliable, simple, and robust method for trace levels of UO22+ detection in environmental samples.
研究领域:
生物化学
·
环境科学
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产品关联
DOI
PubMed
高影响因子
BIOSENSORS & BIOELECTRONICS
·
2025
·
IF 10.5
Jiwei Wang,Chuankun Yang,Ran Xu,Shuo Ma,Ai Chen,Yuming Yao,Fengfeng Zhao,Guoqiu Wu
This method pioneers a synergistic strategy combining ‘extended-strand unmasking with freeze-drying pre-packaging’. By designing unequal-length double-strand replacement strands that competitively bind to regions flanking the target site, it forces the unfolding of hairpin/cloverleaf conformations, thereby physically exposing concealed targets. Concurrently, a five-step lyophilization process (pre-folding-anchoring-sealing-encapsulation-freezing) employs graphene oxide (GO) π-π stacking to suppress >90 % of DNA breathing effects. Combining 0.02 % (w/v) low-melting-point agarose achieves in situ encapsulation and probe conformation fixation, forming microchannels for water molecules. The final lyophilized product forms a porous solid matrix (integrable into nitrocellulose test strips or 2-mm diameter lyophilized spheres), enabling stable probe storage at ambient temperature. This technology overcomes the ‘impossible triangle’ challenge in molecular diagnostics: achieving high room-temperature stability (>6 months, compared to <7 days for traditional liquid-phase CHA), ultra-high sensitivity (0.01 pM), and a simple, rapid testing process. This solution provides a “sample-in, result-out” point-of-care testing approach for home health management (no cold chain required), primary care clinics (no equipment dependency), and pandemic emergency response.
✓ 已验证
产品关联
DOI
PubMed
高影响因子
SENSORS AND ACTUATORS B-CHEMICAL
·
2020
·
IF 7.1
Meng Xiao,Weihua Zhao,Hongbo Li,Jiamei Pu,Mingbin Liu,Suqin Wang,Jianguo Xu,Ruqin Yu
A label-free DNA sensor is proposed to detect the acute leukemia gene pax-5a, in which the system only relies on a functional hairpin probe (HP) containing a G-quadruplex sequence and the cleavage of exonuclease Ⅲ (Exo Ⅲ). Non-single-stranded fragments in the HP structure can also be digested, including double-stranded stems and G-quadruplex fragments, by Exo Ⅲ at a certain concentration. In the presence of pax-5a, the HP sequence can be cyclically opened through the shearing action of Exo Ⅲ, causing a large number of G-quadruplex fragments to be released. In the absence of pax-5a, the G-quadruplex fragment is digested by Exo Ⅲ. The G-quadruplex-hemin DNAzyme can catalyze the oxidation of 3, 3′, 5, 5′-tetramethylbenzidine by H 2 O 2 to generate a detectable visible absorption signal, thereby achieving facile and visual detection of acute leukemia gene pax-5a. The detection limit of the label-free DNA sensor that we proposed is as low as 25 fM in the pax-5a concentration range of 25 fM to 25 nM. In addition, the sensor has good discrimination ability of non-target genes and excellent detection ability in complex matrices. Therefore, the DNA sensor shows great potential in biological analysis and clinical diagnosis.
研究领域:
分子生物学
·
基因工程
·
癌症诊断
·
生物化学
✓ 已验证
DOI
高影响因子
ANALYST
·
2019
·Vol. 144
·期 9
·3023-3029
·
IF 4.019
Qianqian Pei,Xiaolei Song,Su Liu,Jingfeng Wang,Xueqi Leng,Xuejun Cui,Jinghua Yu,Yu Wang,Jiadong Huang
A facile signal-on electrochemical DNA biosensor has been developed for ultrasensitive detection of pathogenic bacteria using an Exo III-assisted autonomous multiple-cycle amplification strategy. The strategy relies on pathogens and aptamer binding-initiated release of a trigger, which combines with the 3′-protruding terminus of the hairpin probe 1, leading to the formation of double-stranded DNA with a blunt 3′ terminus which starts the Exo III-assisted multiple signal amplification reaction. In addition, hairpin probe 2 labeled with an electroactive reporter at the middle of the loop region is ingeniously designed to contain a short hairpin-embedded segment, which can fold into a hairpin structure via an Exo III-assisted cleavage reaction, thus bringing the redox molecule in proximity to the electrode surface for “signal-on” sensing. Under optimal conditions, this biosensor exhibits a very low detection limit as low as 8 cfu mL−1 and a wide linear range from 1.0 × 101 to 1.0 × 107 cfu mL−1 of target pathogenic bacteria. As far as we know, this is the first time that the Exo III-assisted autonomous multiple-cycle amplification strategy has been used for signal-on electrochemical sensing of pathogenic bacteria. In addition, the proposed sensor can also be used for highly sensitive detection of other targets by changing the aptamer sequence, such as nucleic acids, proteins and small molecules. Therefore, the proposed signal-on electrochemical sensing strategy might provide a simple and practical new platform for detection of pathogenic bacteria and related biological analysis, food safety inspection and environmental monitoring.
研究领域:
生物化学
·
电化学
·
分析化学
·
微生物学
✓ 已验证
产品关联
DOI
PubMed
高影响因子
SENSORS AND ACTUATORS B-CHEMICAL
·
2019
·
IF 6.393
Jiaojiao Zhou,Wenjing Wang,Shuojun Li,Axiu Nie,Zhiyong Song,Mohamed F. Foda,Zhicheng Lu,Ling Zhao,Heyou Han
An innovative fluorescent and colorimetric dual-mode strategy based on Y-shaped DNA was proposed for the detection of rabies virus oligonucleotide. The cascade assembly of hairpin probes was initiated by the target, leading to the formation of Y-shaped DNA accompanied by freeing target to catalyze the next assembly process. The fluorescence recovery of the fluorophore FAM was accompanied by opening of hairpin probe. A colorimetric signal was generated by the proximity of glucose oxidase (GOx) and horseradish peroxidase (HRP), which was reflected by enhanced catalytic capability to TMB. Using this method, target DNA can be detected with a wide linear range from 0.1 to 6 nM with a detection limit of 15 pM, and from 0.5 to 60 nM with a detection limit of 60 pM for fluorescent and colorimetric readout, respectively. Moreover, the dual-mode detection strategy exhibited excellent selectivity towards the target compared with interference oligonucleotides. In addition, recoveries ranging from 96.0% to 106% with RSD values varying from 3.2% to 5.4%, and from 91.0% to 102% with RSD values varying from 3.3% to 7.1% for fluorescent and colorimetric studies, respectively, were obtained in diluted cerebrospinal fluid samples. The proposed dual-mode sensor is easy to operate with excellent detection performance. It would be superior to single mode detection and more practicable for quantitative detection of nucleic acid-related biomarkers due to the self-correction capability.
研究领域:
生物化学
·
分子生物学
·
分析化学
·
病毒学
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产品关联
DOI
高影响因子
ANALYTICAL AND BIOANALYTICAL CHEMISTRY
·
2022
·Vol. 414
·期 20
·6157-6166
·
IF 4.478
Liang Yuling,Yang Huihui,Yin Wen,Zhang Yanfei,Xu Yuzhi,Liu Si-Yang,Dai Zong,Zou Xiaoyong
Long-term and continuous monitoring of the microRNA (miRNA) expression in living cells is essential in biomedical research, but it is currently limited by fast consumption and easy digestion of probes in the intracellular environment. Herein, we report polydopamine-modified gold nanoparticles (AuNPs@PDA) as protective and efficient nanocarriers for DNA hairpin probes (hpDNA), achieving long-term monitoring (48 h) of the miRNA (let-7a) levels in living cells after drug treatments. This method enabled excellent sensitivity and high selectivity toward let-7a with a limit of detection of 0.51 nM ( n = 3) and a linear range from 1 to 100 nM. More importantly, AuNPs@PDA can not only efficiently improve the loading of hpDNA on each nanoparticle, but also effectively protect hpDNA from hydrolysis in the cell microenvironment, finally realizing the continuous monitoring of let-7a in living cells for 48 h. This simple method would be of great significance for drug screening and precision medicine. Graphical abstract
研究领域:
生物医学工程
·
纳米技术
·
分子生物学
·
药物发现
✓ 已验证
产品关联
DOI
PubMed
高影响因子
JOURNAL OF CHROMATOGRAPHY A
·
2019
·
IF 3.858
Xixue Chen,Feng Hong,Weilin Zhang,Dazhen Wu,Tianhua Li,Futao Hu,Ning Gan,Jianyuan Lin,Qiqin Wang
It is crucially important to rapidly, simultaneously, and sensitively determine trace amounts of heavy metal ions in complex samples. Herein, a stirring bar modified with two kinds of encoded hairpin DNA probes (H 0 and H 0 ′) was used in a multiplexed strategy allowing for specific extraction of Hg 2+ and Ag + coupled to microchip electrophoresis (MCE) separation and LED induced fluorescence (LIF) detection. The extraction step utilizes stir bars, which are functionalized with designed hairpin DNA probes (H 0 with T T and H 0 ′ with C C mismatches in stems). This allows the specific capture of Hg 2+ and Ag + through C Ag + C and T Hg 2+ T interactions. These complexes are then enzymatically degraded by the action of exonuclease III (Exo III). The ions released during this enzymatic reaction can initiate a new cycle of interactions with hairpin structures and enzymatic reactions and so on. This cyclic step is specific to the presence of Hg 2+ and Ag + and represents the first round of amplification of the presence of the selected ions. The resulting single strand DNAs on the stirring bars after enzymatic degradation were used in the second step as primers to trigger the catalytic hairpin assembly (CHA) in the presence of a couple of hairpin structures in solution. Such a reaction allows producing duplexes that can be monitored by MCE-LIF. The fluorescence intensity of CHA products (I P ) increased and that of hairpin DNAs (I R ) decreased with the increase of target concentrations. The signal ratios (I P /I R and I P ′/I R ′) consisted of targets. The assay was employed for Hg 2+ and Ag + detection in several mediums including water, milk, and fish samples with complex matrices. The results showed that the assay could avoid matrix interference to increase the sensitivity. Therefore, the multiplexed assay was ideal to simultaneously and quickly detect metal ions in complex samples.
研究领域:
分析化学
·
环境科学
·
生物技术
✓ 已验证
产品关联
DOI
PubMed
高影响因子
BIOSENSORS & BIOELECTRONICS
·
2022
·
IF 12.545
Ying Peng,Bing Lu,Ying Deng,Nana Yang,Genxi Li
Accurate and sensitive assay of specific circulating tumor cells (CTCs) is of importance for the diagnosis, treatment, and metastasis monitoring of cancer. Herein, we have proposed a dual-recognition-controlled electrochemical biosensor in this work for the detection of specific CTCs. To this sensor, two aptamer hairpin probes are designed to be able to separately bind to two adjacent proteins on the cell membrane to activate the associative toehold for strand displacement reaction, which will then trigger a dimer-like rolling cycle amplification reaction, and finally produce significantly amplified electrochemical signals for sensitive quantification of target CTCs. In our design, only the case that the two proteins are simultaneously expressed on the cell membrane can result in obvious signal responses, which may greatly improve the accuracy of CTCs analysis. The proposed biosensor can possess excellent selectivity to distinguish target cells from different cancer cells. Moreover, the combination of rolling cycle amplification and DNA nanostructure capture probes can effectively lower the detection limit to 3 cells mL −1 . Notably, our biosensor can be applied to the assay of the target CTCs in the complex whole blood matrixes, verifying its strong stability and anti-interference. Thus, the as-proposed dual-recognition-controlled electrochemical biosensor may exhibit great promise in clinical cancer diagnosis and personalized medicine.
研究领域:
癌症研究
·
生物工程
·
电化学
·
分子生物学
·
临床诊断
✓ 已验证
DOI
PubMed
高影响因子
危险品化学品经营许可证（带存储）