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ANALYTICA CHIMICA ACTA · 2023 · IF 6.2

DNA tetrahedron-besieged primer and DNAzyme-activated programmatic RCA for low-background electrochemical detection of ochratoxin A

Qingxin Zhang,Manru Zhang,Zhiqiang Guo,Jingjing Li,Zhixue Zhu,Yu Wang,Su Liu,Jiadong Huang,Jinghua Yu

Ochratoxin A (OTA) is the most toxic class of ochratoxins and has become a major threat to the environment, humans and animals. Therefore, research on the methods for its detection is also more urgent. Herein, we propose a low-background electrochemical biosensor based on a DNA tetrahedron-besieged primer and a DNAzyme-activated programmatic rolling circle amplification (RCA) that can be ultimately utilized for OTA detection in wine samples. Low-background detection can be achieved using the besieged primer via sequenced assembly of DNA tetrahedral nanostructures so that non-specific extensions of primer can be avoided. The target OTA-mediated DNAzyme activation initiates the programmatic RCA. Additionally, the catalytic property of silver nanoclusters (AgNCs) is integrated with the electrochemical assay to achieve high sensitivity for OTA detection. Benefiting from the aforementioned processes, a low-background, and highly sensitive electrochemical biosensor has been successfully constructed. This design is capable of detecting OTA at concentrations from 1 pg/mL to 10 ng/mL, and its lowest concentration limit is 0.773 pg/mL. Simultaneously, its validation in the detection of actual samples reveals that the proposed electrochemical biosensor has a lot of potential in food safety and environmental detection.

研究领域: 生物传感 · 食品安全 · 环境监测
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BIOELECTROCHEMISTRY · 2024 · IF 4.8

Low background catalytic redox recycling coupled with hybridization chain reaction amplification for highly sensitive electrochemical aptamer luteinizing hormone assay

Yuan Ming,Yujie Liu,Daxiu Li,Bingying Jiang,Yun Xiang,Ruo Yuan

The concentration variation of luteinizing hormone (LH) regulates the cell cycle of oocyte meiosis and significantly affect the whole reproductive cycle. Sensitively quantifying the LH biomarker therefore plays an important role for reproductive disease diagnosis. By coupling a new low background catalytic redox recycling strategy with hybridization chain reaction (HCR), we propose a highly sensitive bio-electrochemical aptamer LH sensing method. LH analyte molecules bind aptamer strands in duplex DNAs to liberate ssDNAs, which trigger HCR generation of [Ru(NH 3 ) 6 ]Cl 3 (RuHex)-modified dsDNA polymers on sensor electrode. Subsequent electrochemical redox recycling of RuHex mediated by K 3 [Fe(CN) 6 ] thus exhibits greatly magnified currents for ultrasensitive LH assay. The synergistic integration of HCR signal amplification with low background redox recycling leads to highly enhanced signal-to-noise ratio and sensitivity for detecting LH down to 6.03 pM. In addition, LH sensing in diluted human serums has been tested and verified, making such sensor a robust detection platform for monitoring diverse biomarkers at low levels for early diagnosing diseases.

CAS / MDL: 13746-66-2 14282-91-8
研究领域: 生物化学 · 医学诊断 · 分析化学
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ANALYTICA CHIMICA ACTA · 2025 · IF 6

Low-background and label-free loop-mediated isothermal amplification for rapid detection of foodborne pathogens using amplicon-templated copper nanoclusters

Wenyu Du,Shuanglong Wang,Yining Luo,Xiaoyu Tong,Yin Chen,Zhipeng Fang,Junzhuo Shan,Daofeng Liu,Haiyan Ni,Weihua Lai,Shan Shan

Background Loop-mediated isothermal amplification (LAMP) is a simple, low-cost, and instrument-free assay, which is highly adaptable for the detection of foodborne pathogens. However, current methods have a high background, making it difficult to achieve sensitive on-site detection. Results Herein, LAMP-CuNCs assay with a low background signal was established. LAMP amplicons were used as the templates for the synthesis of copper nanoclusters (CuNCs), and LAMP results were determined by the fluorescence intensity of the amplicons-CuNCs. The stability of LAMP amplicons templated CuNCs was improved by the addition of 1.5 g/mL fructose, which effectively prolonged the fluorescence lifetime. The detection limit of Salmonella DNA was 756 pg/μL, and the sensitivity and specificity of this method were 12/12 and 12/12, respectively. This method could detect Salmonella at a minimum concentration of 1910 CFU/mL in spiked salmon samples. Significance The study provided a method that can quickly obtain results within 16 min for detection of genomic DNA and visual inspection under a 365 nm UV flashlight. In addition, this method could also be used to detect Listeria monocytogenes , Vibrio parahaemolyticus , and Escherichia coli .

CAS / MDL: 7647-14-5
研究领域: 食品科学 · 分子生物学
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Journal of Materials Chemistry B · 2022 ·Vol. 10 ·期 32 ·6107-6117 · IF 7.571

A DNA–Cu nanocluster and exonuclease I integrated label-free reporting system for CRISPR/Cas12a-based SARS-CoV-2 detection with minimized background signals

Siying Xie,Cheng Qin,Furong Zhao,Zhaoyang Shang,Pei Wang,Muhammad Sohail,Xing Zhang,Bingzhi Li

CRISPR-driven biosensing is developing rapidly, but current studies mostly adopt dye-labeled ssDNA as the signal reporter, which is costly and unstable. Herein, we developed a label-free and low-background reporter for CRISPR/Cas12a signaling by integrating DNA-templated copper nanoclusters (DNA–CuNCs) and exonuclease I (EXO I). The template of the DNA–CuNCs was rationally designed as a ds-/ss-DNA hybrid, ensuring that after a quick and nonpersistent cut of Cas12a, a majority of the template can be digested by EXO I. Based on this novel reporter, a biosensor termed CRISPR-CNS (cost-effective, nimble, and sensitive copper nanocluster sensor integrating CRISPR) was developed. Due to the high signal-to-background ratio of our proposed reporter, CRISPR-CNS shows excellent performances for nucleic acid detection, yielding a detection limit of 20 copies for SARS-CoV-2 RNA. Considering its facile synthesis, robust fluorescence, effective cost, and good sensitivity, this combination shall serve as a highly potential output for CRISPR-based point-of-care testing.

研究领域: 生物技术 · 分子诊断 · 纳米技术 · 病毒学
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ANALYTICA CHIMICA ACTA · 2017 · IF 4.95

Low-background and visual detection of antibiotic based on target-activated colorimetric split peroxidase DNAzyme coupled with dual nicking enzyme signal amplification

Xuejun Cui,Rongguo Li,Xiaofei Liu,Jingfeng Wang,Xueqi Leng,Xiaolei Song,Qianqian Pei,Yu Wang,Su Liu,Jiadong Huang

Herein, we have reported the development of a simple, rapid, and low cost colorimetric method for the detection of antibiotic based on target-activated split peroxidase DNAzyme coupled with dual nicking enzyme signal amplification (NESA). To lower background signal in G-quadruplex DNAzyme-based detection, the two split G-rich parts are caged into two different hairpin probes, respectively, preventing the two parts from assembling into the G-quadruplex structure. By the combination of restriction endonuclease-assisted cleavage reaction with the spilt G-quadruplex probes, target-modulated release of the two split G-rich parts is achieved, affording high specificity of antibiotic detection. Our strategy features with several aspects. First, the less background signal produced by the self-assembly of G-quadruplex in the absence of target is effectively eliminated owing to the pre-blocking of the two split G-rich parts. Second, dual NESA coupled G-quadruplex DNAzyme amplification strategy is integrated with colorimetric assay of antibiotic, which significantly improves the detection sensitivity. Third, peroxidase-mimicking DNAzyme is used as biocatalyst in our reaction system, which can catalyze the oxidation of 2,2' - azino - bis (3 - ethylbenzothiozoline - 6 - sulfonic acid) (ABTS 2− ) mediated by H 2 O 2 to generate the colored radical anion (ABTS •- ), allowing to low cost and visual detection of antibiotic by the naked eye. Under optimized conditions, the results revealed the proposed biosensor exhibits excellent specificity and sensitivity toward kanamycin with a detection limit as low as 14.7 pM. Hence, the target-activated split G-quadruplex DNAzyme and dual NESA-based strategy provides a useful and practical platform for antibiotic residues determination and other analytes detection in bio-analysis.

研究领域: 生物化学 · 分析化学 · 生物技术
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TALANTA · 2017 · IF 4.162

A novel BODIPY-based fluorescent probe for selective detection of hydrogen sulfide in living cells and tissues

Yan Ji,Li-Jun Xia,Liu Chen,Xiao-Feng Guo,Hong Wang,Hui-Juan Zhang

Hydrogen sulfide (H 2 S) is a critical biological messenger in numerous physiological and pathophysiological processes. Small-molecule fluorescent probes combined with fluorescent microscopy have been developed for the sensitive detection of H 2 S. Here, we designed and synthesized a long-wavelength BODIPY-based probe (TMSDNPOB) based on the thiolysis of dinitrophenyl ether according to photo-induced electron transfer theory and the results of computational calculation. 4,4-Difluoro-8-phenyl-1,5,7-trimethyl-3-(4-(2,4-dinitrophenoxy)styryl- 4-bora-3a,4a-diaza-s-indacene (TMSDNPOB) is nearly non-fluorescent, but the reaction product (TMSHOB) emits strong red fluorescence at 592   nm when excited at 574   nm. The long wavelengths of the designed probe indicate low background interference from biological matrix and little photo damage on fluorescence imaging of H 2 S. With the advantages of the turn-on probe for H 2 S including high sensitivity, high selectivity , good biocompatibility and low toxicity, the probe has been used for imaging H 2 S in living cells and liver tissues.

研究领域: 化学 · 生物化学 · 分子生物学 · 荧光成像 · 生物信号
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BIOSENSORS & BIOELECTRONICS · 2017 · IF 7.78

Cross-catalytic hairpin assembly-based exponential signal amplification for CRET assay with low background noise

Shuzhen Yue,Tingting Zhao,Hongjie Qi,Yongcun Yan,Sai Bi

A toehold-mediated strand displacement (TMSD)-based cross-catalytic hairpin assembly (C-CHA) is demonstrated in this study, achieving exponential amplification of nucleic acids . Functionally, this system consists of four hairpins (H1, H2, H3 and H4) and one single-stranded initiator (I). Upon the introduction of I, the first CHA reaction (CHA1) is triggered, leading to the self-assembly of hybrid H1·H2 that then initiates the second CHA reaction (CHA2) to obtain the hybrid H3·H4. Since the single-stranded region in H3·H4 is identical to I, a new CHA1 is initiated, which thus achieves cross operation of CHA1 and CHA2 and exponential growth kinetics. Interestingly, because the C-CHA performs in a cascade manner, this system can be considered as multi-level molecular logic circuits with feedback mechanism. Moreover, through incorporating G-quadruplex subunits and fluorescein isothiocyanate (FITC) in the product of H1·H2, this C-CHA is readily utilized to fabricate a chemiluminescence resonance energy transfer (CRET) biosensing platform, achieving sensitive and selective detection of DNA and microRNA in real samples. Since the high background signal induced by FITC in the absence of initiator is greatly reduced through labeling quencher in H1, the signal-to-noise ratio and detection sensitivity are improved significantly. Therefore, our proposed C-CHA protocol holds a great potential for further applications in not only building complex autonomous systems but also the development of biosensing platforms and DNA nanotechnology.

研究领域: 分子生物学 · 生物化学 · 分析化学 · 纳米技术 · 生物医学工程
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FOOD CONTROL · 2023 · IF 6

Cascade amplifying aptasensor for positively correlated detecting OTA: Based on DNase I-assisted cyclic enzyme digestion and AgNPs@gel-enhanced fluorescence

Chenfei Han,Yu Xiao,Zhenjiang Liu,Daolin Du,Ming Li

To improve the performance of aptasensor, the coupling of novel material and strategy had been the hottest research with the promising prospects. In this study, a cascade amplifying aptasensor (CA-aptasensor), which had coupled the DNase I-assisted cyclic enzyme digestion and AgNPs@gel-enhanced fluorescence, had reflected the desired performance for positively correlated detecting OTA . For this purpose, the GO@PVP was used to adsorb and quench the free FAM-Apt, while the complex of OTA and FAM-Apt was digested by the DNase I. The released OTA allowed the free FAM-Apt to separate from the GO@PVP cyclically, thus the OTA level was amplified preliminarily. The AgNPs@gel could combine with the released FAM to form the metal enhanced fluorescence (MEF) effect, resulting in the cascade signal amplification. The OTA was detected at pg mL −1 level with an ultrasensitive LOD of 3.33 pg mL −1 and IC 50 of 13.6 pg mL −1 , which improved 90-fold and 728-fold than that of ELASA for same aptamer. The CA-aptasensor indicated high specificity, reliability and practicability. Meanwhile, the CA-aptasensor exhibited more highlights, including the efficient homogeneous system, low background signal and standardized aptamer recognizer. This study might provide a new approach for improving the performance of aptasensor, which could also guide the signaling cascade and positively correlated detection for other risk factors.

研究领域: 分析化学 · 生物传感器 · 纳米技术 · 食品安全
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TALANTA · 2021 · IF 6.057

Label-free photoelectrochemical biosensor for alpha-fetoprotein detection based on Au/CsxWO3 heterogeneous films

Jing Li,Dali Liu,Donglei Zhou,Long Shao,Xu Chen,Hongwei Song

In this work, a novel label-free photoelectrochemical (PEC) biosensor based on Au/Cs x WO 3 heterogeneous films with low background noise and excellent sensitive detection of alpha-fetoprotein (AFP) was proposed. We prepared Au/Cs x WO 3 heterogeneous films by different methods including template removal and spin coating . The surface plasmon resonances (SPR) of Au film and Cs x WO 3 semiconductor nanocrystals largely improved the photoelectrochemical properties of the electrodes and enhanced the photocurrent . Under the optimal conditions, the prepared PEC biosensor showed a linear response between photocurrent variation and the logarithm of AFP concentration in the range from 0.01 ng/mL to 500 ng/mL with a low detection limit of 7 pg/mL. In addition, the proposed PEC biosensor had excellent specificity, repeatability, long-term stability and showed satisfactory results in the analysis of human serum samples, which would have great prospect in clinical application and biological analysis in the future.

CAS / MDL: 10294-29-8
研究领域: 生物传感器 · 光电化学 · 纳米材料 · 生物医学工程 · 分析化学
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TALANTA · 2021 · IF 6.057

A novel fluorescent strategy based on double modifications of metal organic framework material CAU-10-NH2 for low background and high sensitivity determination of H2S

Wenpeng Gong,Li Zhang,Ying Yu,Bixia Lin,Yumin Wang,Manli Guo,Yujuan Cao

Hydrogen sulfide is typical metabolic marker and environmental pollutant which is worthwhile to determine. Herein, a low background and high sensitivity fluorescent strategy based on double modifications of metal organic framework material CAU-10-NH 2 is proposed for the determination of hydrogen sulfide. Firstly, a functional monomer 3,5-diaminobenzoic acid is employed to modify on the CAU-10-NH 2 , the product CAU-10-NH-dAba has strong fluorescent performance at 412 nm under an excitation wavelength of 320 nm. Subsequently, it is further modified by the azide group to form CAU-10-NH-dAba-N 3 . This azidation inhibits the fluorescent signal. However, in the presence of hydrogen sulfide, the azide group is specifically reduced to amidogen, and results in the recovery of the fluorescence. The CAU-10-NH-DABA-N 3 was characterized by solid state NMR , XPS, fluorescence, IR, XRD, SEM and specific surface area. After the optimization of pH value, temperature and interaction time, the detection results of hydrogen sulfide demonstrate the linear range of this strategy is from 20 to 140 nM with a detection limit of 1.51 nM, which is significantly better than that of the CAU-10-NH 2 merely modified by 3,5-dinitrobenzoic acid. Meanwhile, the satisfactory assay results of hydrogen sulfide in serum sample and Pearl river water suggest a potential application prospect of this strategy in clinical diagnosis and environment monitoring.

研究领域: 分析化学 · 环境科学 · 材料科学 · 生物化学
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