JOURNAL OF AGRICULTURAL AND FOOD CHEMISTRY
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2021
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IF 5.279
Yiming Xie,Haiyan Wang,Xinyue Yuwen,Guosong Lai
Herein, we combine the exonuclease III (Exo III)-catalyzed release of a Zn2+-dependent ligation DNAzyme with the DNAzyme-driven strand displacement reaction (SDR) to develop a novel homogeneous colorimetric bioassay method for kanamycin (Kana) antibiotic detection. Upon the biorecognition reaction between Kana and a designed hairpin DNA, the DNAzyme-containing strand can be catalytically released by Exo III. Then, this DNAzyme will catalyze the ligation of two oligonucleotides to cause a SDR and the aggregation of gold nanoparticles (Au NPs) labeled by two linker DNA strands. Due to the aggregation of Au NPs for colorimetric signal transduction and the Exo III and SDR-assisted dual signal amplification, this method shows a wide linear range of 5 orders of magnitude and a very low detection limit down to 8.1 fg mL–1. Together with its excellent selectivity, repeatability, reliability, and convenient manipulation, the proposed method shows a great potential for the food quality monitoring application.
研究领域:
分析化学
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生物化学
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纳米技术
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食品科学
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高影响因子
ANALYTICA CHIMICA ACTA
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2024
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IF 5.7
Zaiyong Zhang,Jinglong Li,Chunlin Chen,Yuwei Tong,Dehui Liu,Cuizhi Li,Huan Lu,Li Huang,Wanling Feng,Xiaoting Sun
Brain natriuretic peptide (BNP) is considered to be an important biomarker of heart failure (HF) attracting attention. However, its low concentration and short half-life in blood lead to a low-sensitivity detection of BNP, which is a challenge that has to be overcome. In this work, we propose a highly specific, highly sensitive T7 RNA polymerase-assisted clustered regularly interspaced short palindromic repeats (CRISPR)/Cas13a system to detect BNP via an electrochemiluminescence (ECL) sensing platform and incorporate Exo III-hairpin and dumbbell-shaped hybridization chain reaction (HCR) technologies. In this detection scheme, the ECL sensing platform possesses low background signal and high sensitivity. Firstly, the T7 promoter-initiated T7 RNA polymerase acts as a signal amplification technique to generate large amounts of RNAs that can activate CRISPR/Cas13a activity. Secondly, CRISPR/Cas13a is able to trans -cleave the surrounding trigger strand to produce DNA1. Thirdly, DNA1 is involved in the co-amplification reaction of Exo III and hairpin DNA, which subsequently triggers a dumbbell-shaped HCR technology. Eventually, a large number of Ru (II) molecules are inserted into the interstitial space of the dumbbell-shaped HCR to generate a strong ECL signal. The CRISPR/Cas13a possesses outstanding specificity for a single base and increased sensitivity. The tightly conformed dumbbell-shaped HCR provides higher sensitivity than the traditional linear HCR amplification technique. Ultimately, the clever combination of several amplification reactions enables the limit of detection (LOD) as low as 3.2 fg/mL. It showed promise for clinical sample testing, with recovery rates ranging from 98.4% to 103% in 5% human serum samples. This detection method offered a valuable tool for early HF detection, emphasizing the synergy of amplification strategies and specificity conferred by CRISPR/Cas13a technology.
研究领域:
生物技术
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分子生物学
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分析化学
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临床诊断
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高影响因子
MICROCHEMICAL JOURNAL
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2026
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IF 5.2
Beili Wang,Wei Gao,Liang Yang
An Exo-III-triggered target-reconstructed DNAzyme walker enables ultrasensitive multiplex miRNA detection. • Cascade amplification integrates Exo-III recycling, polymerase/endonuclease cycling, and DNAzyme walking. • Orthogonal Pt, Ir, Rh tags allow simultaneous ICP-MS quantification of three miRNAs without spectral cross-talk. • Detection limits reach 0.15 fM (miR-21), 0.32 fM (miR-1246), and 0.26 fM (miR-20a) with excellent linearity. • The method discriminates single-base mismatches and performs well in diluted human serum (92–108% recovery). Cervical cancer remains a leading cause of cancer-related mortality among women, and the prognosis of patients after radiotherapy varies considerably due to differences in tumor radiosensitivity and treatment response. Reliable biomarkers for post-radiotherapy prognosis are therefore critically needed. MicroRNAs (miRNAs) have emerged as promising non-invasive indicators, yet accurate detection is hindered by their low abundance and the lack of multiplexing capability in conventional methods. Herein, we report a highly sensitive and specific platform for simultaneous detection of three cervical cancer radiotherapy-associated miRNAs (miR-21, miR-1246, and miR-20a) by integrating an Exo-III-triggered target-reconstructed DNAzyme walker with inductively coupled plasma mass spectrometry (ICP-MS). The strategy employs a structurally locked DNA walker that is sequentially activated through Exo-III-mediated target recycling and DNA polymerase/endonuclease-assisted cyclic amplification, ultimately unleashing a DNAzyme walker to cleave element-labeled substrates. Using orthogonal platinum group element tags (Pt, Ir, Rh), the released tags are quantified by ICP-MS without spectral cross-talk. The method achieves detection limits as low as 0.15 fM, 0.32 fM, and 0.26 fM for miR-21, miR-1246, and miR-20a, respectively, with excellent single-nucleotide discrimination and negligible cross-reactivity. Successful spike-recovery experiments in diluted human serum validate its practical feasibility. This cascade amplification strategy offers a robust, multiplex, and ultrasensitive tool for non-invasive liquid biopsy-based prognostic assessment of post-radiotherapy cervical cancer, and can be readily adapted for other disease-related miRNA panels. Download: Download high-res image (121KB) Download: Download full-size image
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高影响因子
FOOD CHEMISTRY
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2025
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IF 9.8
Menghui Wang,Ye Tian,Fengjuan Zhao,Zhiguang Suo,Yong Liu,Min Wei
Acrylamide (AA) contamination in food sources is an issue that endangers both human health and food safety worldwide; therefore, developing a highly sensitive detection technique is necessary. Herein, a boron and nitrogen co-doped carbon flower–like composite (B/N C NF) was prepared as a highly efficient quencher for the first time, and its quenching efficiency was found to be approximately 4 fold higher than that of N C. Three hairpin probes and exonuclease III were used to provide a double-cycle amplification system to improve the sensitivity. The sensor had a linear range of 0.01–500 nM, with limit of detection of 1.802 pM. In addition, the sensor successfully realized the detection of AA in actual samples (coffee, potato chips) and quality control samples (potato chips, cookies), and were compared with high performance liquid chromatography-mass spectrometry and high performance liquid chromatography, and the results showed that the sensor had reliable accuracy.
研究领域:
分析化学
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食品科学
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材料科学
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生物化学
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环境科学
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高影响因子
ANALYTICAL LETTERS
·
2019
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IF 1.248
Chunyan Wang,Hui Zhang,Xiaoqing Jiang,Bo Zhou
A sensitive electrochemical biosensor was designed for determination of aflatoxin B1 (AFB1) using a copper-based metal-organic framework (Cu-MOF), which has strong electrochemical activity and exonuclease III (Exo III)-assisted recycling for dual signal amplification. Hairpin DNA (S1) was immobilized on the electrode. The AFB1 was recognized by aptamer DNA (S2) and complementary DNA (S3) was released. The S3 hybridized with the hairpin S1 to form the Exo III hydrolyzed double-stranded DNA, leaving a partial sequence of hairpin DNA (S1′) on the electrode and releasing S3 for the next cycle of the opening and digestion of hairpin S1. The amplified S1′ then was able to combine with more signal probes. Cu-MOF bond gold nanoparticles (AuNPs) by -NH2 were immobilized to capture DNA (S4) to obtain Cu-MOF/AuNPs/S4. This signal probe Cu-MOF/AuNPs/S4 was able to hybridize with the electrode and generate an amplified electrochemical signal. Under the optimized conditions, this electrochemical biosensor for AFB1 exhibited a low detection limit of 6.7 × 10−7 ng/mL at a signal-to-noise equal to 3 and a wide linear range from 10−6 to 1 ng/mL. The biosensor was also used to analyze AFB1-spiked beer sample with recovery values between 96% and 103%. This method has the potential to become a valuable technology for detecting various toxins by the selection of the appropriate aptamer DNA.
研究领域:
分析化学
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生物技术
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食品安全
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环境科学
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高影响因子
ANALYTICAL AND BIOANALYTICAL CHEMISTRY
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2020
·Vol. 412
·期 11
·2487-2494
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IF 3.637
Xue Ning,Wu Shujie,Li Zongbing,Miao Xiangmin
A versatile nanocomposite was simply prepared based upon the electrostatic adsorption of positively charged gold nanoparticles with negatively charged graphene oxide ( nano-gold@GO), and utilized as a novel fluorescence quenching platform for ultrasensitive detection of adenosine triphosphate (ATP). In the designed system, DNA-stabilized Ag nanoclusters (DNA/AgNCs) were used as fluorescent probes, DNA duplex was formed in the presence of ATP, and they can electrostatically adsorb onto the surface of nano-gold@GO to quench the fluorescence signal. Upon the addition of exonuclease III (Exo III), the DNA duplex would be hydrolyzed into DNA fragments and resulted in the recovery of the fluorescence signals due to the diffusion of AgNCs away from nano-gold@GO. Based on these, sensitive detection of ATP was realized with a detection range of 5.0 pM–20 nM. Notably, a good recovery in the range of 94–104% was obtained when detecting ATP in human serum samples, indicating a promising application value in early disease diagnosis.
A functional positively charged nano-gold@graphene oxide was fabricated and utilized as an enhanced fluorescence quenching platform for the detection of ATP, coupled with exonuclease III-assisted signal amplification.
研究领域:
纳米技术
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生物医学工程
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分析化学
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分子生物学
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PubMed
高影响因子
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