MICROCHEMICAL JOURNAL
·
2024
·
IF 4.9
Yizhi Zhou,Qiang Liu
The sensitive and label-free detection of microRNA (miRNA) is crucial for the comprehension of miRNA-related pathological process, such as glioma. Here, we utilized a SYBR Green I (SG) catalytic photosensitization colorimetric reaction in conjunction with target recognition-induced exonuclease-iii (Exo-iii) assisted cascade toehold assembly amplification to achieve highly sensitive and label-free miRNA detection. In this research, the photosensitized generation of 1 O 2 by SG could directly oxidize 3,3′,5,5′-tetramethylbenzidine (TMB) without the need for H 2 O 2 oxidation or enzyme catalysis, which simplifies the experimental procedures and renders the method highly stable. Furthermore, the Exo-iii enhances the target recognition induced catalytic chain assembly, thereby forming cascade signal amplification and providing the method with a significantly higher level of sensitivity than conventional colorimetric miRNA detection methods. Based on this, the proposed method exhibits a low limit of detection of 2.1 fM for miRNA-212 detection. Taking the merit of the high sensitivity, portability, and simplicity, the proposed method can serve as a highly effective nucleic acid detection tool and can facilitate the advancement of biosensors for point-of-care testing (POCT) and clinical disease diagnosis.
研究领域:
生物化学
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分子诊断
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医学检测
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产品关联
DOI
高影响因子
TALANTA
·
2024
·
IF 5.6
Lifeng Zhang,Shihua Luo,Rui Fan,Ruixi Li,Wenbin Li,Siting Chen,Fei Lan,Yitong Zhu,Tingting Ji,Ye Zhang,Ling Li
APE1, an essential enzyme for DNA repair, is overexpressed in various cancers and has been identified as a potential biomarker for cancer diagnosis. However, detecting APE1 at low expression levels in the early stage of cancer presents a significant obstacle. Here, we introduced a novel localized Cas12a-based cascade amplification (LCas12a-CA) method. This method confined both the terminal deoxynucleotidyl transferase and the crRNA/Cas12a complex onto the surfaces of gold nanoparticles (AuNPs). This confinement not only boosts the stability of the multiple enzymes but also induces a substrate channeling effect. As a result, it significantly accelerates the reaction rate and enhances the sensitivity of APE1 detection. Upon the addition of APE1, the AP sites within the APE1 primer can be recognized and cleaved by APE1, exposing the 3′-OH ends. In the presence of LCas12a-CA, polyA sequences are generated at 3′-OH ends with the help of TdT and dATP. The sequences directly enter the Cas12a system, activating the trans -cleavage activity of Cas12a, thereby cutting the reporters on the surface of AuNPs and releasing fluorescence. Our platform demonstrates a detection limit (LOD) as low as 2.51 × 10 −6 U/mL, which is more than 60 times lower than that of free Cas12a-CA. Furthermore, the LCas12a-CA exhibits enhanced resistance ability in extreme environments and has been proven effective for the detection of APE1 in clinical samples. Overall, this work offers a promising platform for robust biosensing in cancer diagnosis and prognosis.
研究领域:
分子生物学
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癌症研究
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生物技术
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产品关联
DOI
PubMed
高影响因子
ANALYTICAL CHEMISTRY
·
2021
·
IF 6.986
Guo Liu,Shan Huang,Xiaochen Liu,Wanzhen Chen,Xin Ma,Shuang Cao,Liping Wang,Lanlan Chen,Huanghao Yang
Transmembrane signal transduction is of profound significance in many biological processes. The dimerization of cell-surface receptors is one prominent mechanism by which signals are transmitted across the membrane and trigger intracellular cascade amplification reactions. Recreating such processes in artificial systems has potential applications in sensing, drug delivery, bioengineering, and providing a new route for a deep understanding of fundamental biological processes. However, it remains a challenge to design artificial signal transduction systems working by the receptor dimerization mechanism in a predictable and smart manner. Here, benefitting from DNA with features of programmability, controllability, and flexible design, we use DNA as a building material to construct an artificial system mimicking dimerization of receptors for signal transduction and cascade amplification. DNA-based membrane-spanning receptor analogues are designed to recognize external signals, which drives two receptors into close spatial proximity to activate DNAzymes inside the cell-mimicking system. The activation of the DNAzyme initiates the catalyzed cleavage of encapsulated substrates and leads to the release of fluorescent second messengers for signal amplification. Such an artificial signal transduction system extends the range of biomimetic DNA-based signaling systems, providing a new avenue to study natural cell signaling processes and artificially regulate biological processes.
研究领域:
合成生物学
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分子生物学
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纳米生物学
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产品关联
DOI
PubMed
高影响因子
ANALYTICA CHIMICA ACTA
·
2022
·
IF 6.911
Yusi Li,Fang Yang,Shunmei Li,Ruo Yuan,Yun Xiang
Proteins play vital roles in regulating a series of living activities and can be potentially considered to be useful indicators for diagnosing various diseases. Here, a target-triggered transcription cascade amplification strategy with the use of the fluorescence switch-on RNA aptamer is constructed for label-free and sensitive platelet-derived growth factor-BB (PDGF-BB) protein biomarker detection. The target molecule of PDGF-BB binds a rationally designed multifunctional hairpin probe to switch its structure and to expose a primer binding region. The primer sequence further binds such a region to initiate the target recycling and lighting-up RNA aptamer transcription cascade cycles to yield tremendous RNA aptamers. And, the fluorescence of the organic dye is substantially enhanced upon binding to these RNA aptamers for realizing highly sensitive and label-free PDGF-BB detection with the detection limit being lowered to 0.8 pM. Moreover, the developed strategy has superior selectivity and exhibits a promising potential to detect PDGF-BB spiked into serums with buffer dilution, which makes this method an attractive sensing system for detecting other biomarkers at trace levels.
研究领域:
生物化学
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分子生物学
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医学诊断
✓ 已验证
DOI
PubMed
高影响因子
BIOSENSORS & BIOELECTRONICS
·
2018
·
IF 8.173
Zhiqiang Chen,Ying Liu,Chen Xin,Jikuan Zhao,Shufeng Liu
Herein, an autocatalytic strand displacement amplification (ASDA) strategy was proposed for the first time, which was further ingeniously coupled with hybridization chain reaction (HCR) event for the isothermal , label-free and multiple amplification toward nucleic acid detection. During the ASDA module, the target recognition opens the immobilized hairpin probe (IP) and initiates the annealing of the auxiliary DNA strand (AS) with the opened IP for the successive polymerization and nicking reaction in the presence of DNA polymerase and nicking endonuclease . This induces the target recycling and generation of a large amount of intermediate DNA sequences, which can be used as target analogy to execute the autocatalytic strand displacement amplification. Simultaneously, the introduced AS strand can propagate the HCR between two hairpins (H1 and H2) to form a linear DNA concatamer with cytosine (C)-rich loop region, which can facilitate the in-situ synthesis of silver nanoclusters (AgNCs) as electrochemical tags for further amplification toward target responses. With current cascade ASDA and HCR strategy, the detection of target DNA could be achieved with a low detection limit of about 0.16 fM and a good selectivity . The developed biosensor also exhibits the distinct advantages of flexibility and simplicity in probe design and biosensor fabrication, and label-free electrochemical detection , thus opens a promising avenue for the detection of nucleic acid with low abundance in bioanalysis and clinical biomedicine .
研究领域:
生物分析化学
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分子生物学
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临床医学
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产品关联
DOI
PubMed
高影响因子
BIOSENSORS & BIOELECTRONICS
·
2018
·
IF 8.173
Jing Lu,Lin Wu,Yufang Hu,Sui Wang,Zhiyong Guo
In this study, a novel electrochemiluminescence (ECL) biosensor for sensitive detection of femtomolar miRNA-141 was constructed on the basis of Faraday cage-type strategy via graphene oxide (GO) and hybridization chain reaction (HCR)-assisted cascade amplification. A capture probe (CP) was immobilized on Fe 3 O 4 @SiO 2 @Au nanoparticles as capture unit, which could catch the miRNA-141, and the immobilization of the signal unit (Ru(phen) 3 2+ -HCR/GO) was allowed via nucleic acid hybridization. The prepared biosensor exhibited two advantages for signal amplification : firstly, GO could lap on the electrode surface directly, extending Outer Helmholtz Plane (OHP) of the sensor due to the large surface area and good electronic transport property; secondly, HCR-assisted cascade amplification was designed by anchoring all HCR products on the GO surface, then embedding Ru(phen) 3 2+ as a signal readout pathway. All these signal molecules could take part in electrochemical reactions , thus further enhancing the ECL signal drastically. Therefore, the proposed sensor constructed by integrating HCR with Faraday cage-type strategy displayed an ultrasensitive detection platform for the miRNA-141 with a low detection limit of 0.03 fM. In addition, this proposed biosensor provides a universal platform for analysis of other microRNAs.
研究领域:
生物传感器技术
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分子诊断学
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纳米生物学
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DOI
PubMed
高影响因子
SCIENCE OF THE TOTAL ENVIRONMENT
·
2024
·
IF 8.2
Zongwu Wei,Xuzhe Zhang,Yingzhan Chen,Hongjie Liu,Shaopeng Wang,Man Zhang,Honglin Ma,Kefu Yu,Liwei Wang
Population outbreaks of the crown-of-thorns starfish (COTS) seriously threaten the sustainability of coral reef ecosystems. However, traditional ecological monitoring techniques cannot provide early warning before the outbreaks, thus preventing timely intervention. Therefore, there is an urgent need for a more accurate and faster technology to predict the outbreaks of COTS. In this work, we developed an electrochemical biosensor based on a programmed catalytic hairpin assembly (CHA) and hybridization chain reaction (HCR) cyclic amplification strategy for sensitive and selective detection of COTS environmental DNA (eDNA) in water bodies. This biosensor exhibited excellent electrochemical characteristics, including a low limit of detection (LOD = 18.4 fM), low limit of quantification (LOQ = 41.1 fM), and wide linear range (50 fM – 10 nM). The biosensing technology successfully allowed the detection of COTS eDNA in the aquarium environment, and the results also demonstrated a significant correlation between eDNA concentration and COTS number ( r = 0.990; P < 0.001). The reliability and accuracy of the biosensor results have been further validated through comparison with digital droplet PCR (ddPCR). Moreover, the applicability and accuracy of the biosensor were reconfirmed in field tests at the COTS outbreak site in the South China Sea, which has shown potential application in dynamically monitoring the larvae before the COTS outbreak. Therefore, this efficient electrochemical biosensing technology offers a new solution for on-site monitoring and early warning of the COTS outbreak.
研究领域:
海洋生物学
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环境科学
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分析化学
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产品关联
DOI
PubMed
高影响因子
JOURNAL OF FOOD COMPOSITION AND ANALYSIS
·
2025
·
IF 4.6
Xin Xie,Jingxi Wang,Daxiu Li,Ding Wang
Ochratoxin A (OTA) poses a great threat to food and environmental safety owing to its high toxicity. Herein, we developed a sensitive electrochemical aptasensor that integrates aptamer, enzyme-free DNA circuits, and electrochemical technology for OTA detection. In the presence of target OTA, the aptamer recognition first initiated the EDC circuit to releasing DNA (T2) from a ternary DNA complex (S-O-T2). The released T2 as the trigger to activate the CHA circuit and expose amounts of G-quadruplex sequences on modified electrode surface. Eventually, hemin was embedded in the G-quadruplex structure and generated an amplified electrochemical signal. Under optimal conditions, this developed biosensor is capable of detecting OTA ranging from 1 pg/mL to 10 ng/mL with a low detection limit of 0.135 pg/mL, which could be contributed to the design of target-triggered EDC and CHA cascade amplification. Additionally, the proposed biosensor was applied to analyze OTA in corn samples with satisfactory results, demonstrating that the proposed method has great potential in food safety and environmental monitoring.
研究领域:
生物化学
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分析化学
·
食品科学
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产品关联
DOI
高影响因子
BIOSENSORS & BIOELECTRONICS
·
2024
·
IF 10.7
Fang Zhu,Hu Zhang,Ruoxi Wu,Yanyan Lu,Jin Wang,Ravikumar A,Tamil Selvan G,Nuanfei Zhu,Zhen Zhang,Jun Tang
Aflatoxin B1 (AFB1) is considered as a serious carcinogenic mycotoxin that was widely detected in grains and foods, and its sensitive analysis is of key importance to avoid the health threats for consumers. In this study, a dual-signal aptasensor based on cascade of entropy-driven strand displacement reaction (ESDR) and linear rolling circle amplification (LRCA) was fabricated for ultrasensitive determination of AFB1. At the sensing system, the complementary strand would be released after the aptamer combined with AFB1, which will bring about the functional domains exposed, triggering the subsequent ESDR. Meanwhile, the two strands that were outputted by ESDR would incur the downstream LRCA reaction to produce a pair of long strands to assist in the generation of fluorescence and absorbance signals. Under the optimized conditions, the proposed aptasensor could achieve excellent sensitivity (limit of detection, 0.427 pg/mL) with satisfactory accuracy (recoveries, 92.8–107.9 %; RSD, 2.4–5.0 %), mainly ascribed to the cascade amplification. Importantly, owing to the flexibility design of nucleic acid primer, this analytical method can be applied in monitoring various hazardous substances according to the specific requirements. Our strategy provides some novel insights at signal amplification for rapid detection of AFB1 and other targets.
研究领域:
分析化学
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食品安全
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分子生物学
✓ 已验证
产品关联
DOI
PubMed
高影响因子
SENSORS AND ACTUATORS B-CHEMICAL
·
2018
·
IF 5.667
Wei Li,Jiewei Sun,Huijuan Wang,Lei Wang,Wei Jiang
Herein, a multifunctional aptamer probe mediated cascade amplification strategy has been constructed for sensitive and label-free detection of adenosine. First, the multifunctional aptamer probe was designed with an aptamer for adenosine identification and a template for cascade amplification. These two regions could partially hybridize with each other and formed a stem-loop structure. Thus, in the absence of adenosine, the probe could be inhibited by itself and the interference signals could be reduced effectively. In the presence of adenosine, the probe identified the adenosine, resulting in the conformational transformation and the release of the template. Then, the 3′-end of the probe-adenosine complex acted as primer to trigger the cascade amplification of strand displacement amplification (SDA) and catalytic hairpin assembly (CHA), yielding a lot of G-quadruplex. Finally, by inserting N-methy mesoporphyrin IX (NMM), an enhanced and label-free fluorescence signal was achieved. This strategy suggested a high sensitivity with a detection limit of 1.3 × 10 −7 mol/L that attributes to the low interference signals of the probe and the amplification capability of the cascade amplification. Moreover, adenosine analysis in human urines was performed, indicating that this method was reliable and could be further used in the early clinical diagnosis and medical research.
研究领域:
生物化学
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分子诊断
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医学研究
✓ 已验证
产品关联
DOI
高影响因子
NEW JOURNAL OF CHEMISTRY
·
2017
·Vol. 41
·期 15
·7613-7620
·
IF 3.269
Qianqian Pei,Yu Wang,Su Liu,Yifei Qin,Xueqi Leng,Xuejun Cui,Jiadong Huang
In this study, a novel electrochemical biosensor based on exonuclease III (Exo III)-aided autonomous cascade signal amplification for ultrasensitive and highly specific detection of Salmonella typhimurium (S. typhimurium) has been reported. Herein, two duplex DNA probes were ingeniously designed. An Ap–Pr duplex probe was constructed as a recognition element by hybridizing an S. typhimurium aptamer (Ap) with a primer (Pr). A P–S duplex probe has been designed to contain a protruding DNA fragment at 5′-termini that has an identical sequence with Pr (the secondary Pr), a complementary duplex DNA that is used for blocking a signal-triggered probe (S), and an overhung nucleic acid segment at 3′-end that can anneal with Pr and form double-stranded DNA with a blunt 3′-terminus, thus initiating Exo III-aided amplification reaction. Upon sensing of the target S. typhimurium, the specific recognition of S. typhimurium and aptamer releases Pr, which anneals with the P–S duplex and triggers the Exo III cleavage reaction, accompanied by release of Pr, S, and secondary Pr. After this, the released Pr and secondary Pr anneal with other P–S duplex, which in turn triggers the Exo III-assisted cycling cleavage. Thus, large amounts of S are liberated, which hybrids with S* (DNA probe complementary to S); thereby, 5′-methylene blue (MB)-labelled DNA probe (MBP) folds into a hairpin structure with 5′-terminus close to electrode surface for efficient electronic transfer. Moreover, the S–S* hybrid duplex is specifically hydrolyzed by Exo III to liberate S, which accordingly results in the cycling cleavage of S*. Therefore, numerous MBP transform into a hairpin structure with electroactive MB close to the electrode surface; this generates significantly amplified electrochemical signal towards S. typhimurium detection. The present developed strategy exhibited high sensitivity towards S. typhimurium with a detection limit of 2.8 × 10 cfu mL−1. Furthermore, the proposed autonomous cascade signal amplification strategy can be easily extended for detection towards a wide spectrum of analytes including nucleic acid, proteins, and small molecules possessing their corresponding aptamers. Therefore, the developed Exo III-aided autonomous cascade signal amplification strategy might indeed provide a facile and universal tool for bioanalysis and clinical diagnosis.
研究领域:
生物传感技术
·
分子诊断
·
分析化学
✓ 已验证
DOI
高影响因子
CHEMICAL ENGINEERING JOURNAL
·
2023
·
IF 15.1
Ying Zhang,Yunhai Chen,Dianwei Zhang,Huilin Liu,Baoguo Sun
Within this study, a peptide nanodots-bridged metal–organic framework (PNMOF) has been constructed by predictably assembling highly efficient fluorescent dipeptide nanodots into the three-dimensional framework upon coordination of Zn(II) and dihydroxamate linkers. The established system was on behalf of a new class of tripartite hybrid material, i.e., metal ion, organic linker, and peptide nanodot, which integrated the chemical and structural versatility in a modular fashion. Owing to its unique open framework with tripartite construction, PNMOF exhibited a rapid and sensitive response, therefore, the target (e.g., lambda-cyhalothrin, LCY) driven into the tripartite hybrid structure stimulated a cascade amplification effect. LCY can cause significant fluorescence quenching in the PNMOF-based cascade amplification platform due to the charge transfer mechanism, which was confirmed by density functional theory calculations. PNMOF-based cascade amplification platform was employed for fluorescence imaging detection of pyrethroids in solution and solid state assisted by the smartphone. The limit of detection was as low as 0.34 μg/L and the recoveries percentage in real agricultural products was calculated as 88.26–108.17%. This work not only underlies the structural diversity of conventional metal–organic framework but also offers a useful tool in environmental protection and food safety.
研究领域:
分析化学
·
食品安全
·
环境科学
✓ 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）