ANALYTICA CHIMICA ACTA
·
2026
·
IF 6.1
Wen Xu,Yutian Cheng,Kexin Sun,Yue Wu,Yiming Xu,Jiang Ye,Pengfei Li,Haizhen Wu
Background Kanamycin's persistent contamination in agricultural products poses significant human health risks due to its nephrotoxicity and bioaccumulation via the food chain. Existing methods for on-site kanamycin monitoring lack sufficient sensitivity and portability, limiting their utility in field settings. The urgent need for rapid detection technologies remains unmet. This work addresses the critical gap in developing a field-deployable platform for ultrasensitive kanamycin residue screening. Results We engineered a fluorescence biosensor integrating DNAzyme-assisted Cas9 nickase-based amplification reaction (Cas9nAR). Kanamycin binding induces aptamer conformational changes, triggering Cas9nAR-driven cascade amplification that continuously generates DNAzymes. These cleave reporter probes to enable quantitative detection. The system achieved a broad linear range (1 nM – 5 μM) with an ultralow detection limit (0.3 nM), surpassing conventional methods. It exhibited high specificity against interfering antibiotics and delivered consistent recoveries (97% to 103%) in spiked water and milk samples. Following pretreatment, analysis can be completed within 120 min, validating its operational simplicity and robustness for complex matrices. Significance and novelty This work reports the first CRISPR-Cas9 nickase/DNAzyme cascade amplification platform for small-molecule detection, establishing a new paradigm that integrates programmable nucleic acid amplification with catalytic signal turnover. By using Cas9nAR-generated ssDNA as an in situ template for autonomous DNAzyme assembly, the biosensor achieves ultrasensitive, and homogeneous detection of kanamycin-addressing a critical gap in field-deployable antibiotic residue monitoring. The modular design offers a generalizable strategy for translating non-nucleic acid recognition events into amplified fluorescent outputs, with broad implications for point-of-need diagnostics in food safety and environmental analysis.
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DOI
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高影响因子
ANALYST
·
2014
·Vol. 139
·期 17
·4315-4321
·
IF 3.906
Tao Liu,Xiao Zhang,Wenxin Zhu,Wei Liu,Daohong Zhang,Jianlong Wang
The rapid and sensitive detection of Alicyclobacillus acidoterrestris (AA) has become very important due to the frequent occurrence of fruit juice spoilage by AA. In the present study, using guaiacol, both as the metabolic product of AA related to its concentration and as a green colorimetric substrate of G-quadruplex DNAzyme, a novel G-quadruplex DNAzyme-based colorimetric method for a rapid detection of AA has been developed for the first time. Under optimal conditions, AA has been successfully detected in the concentration range of 102–105 cfu mL−1 with a detection limit of 85 cfu mL−1. The recoveries ranging from 71.8% to 115.7% with relative standard deviation from 1.2% to 6.6% in spiked apple and orange juice samples were obtained. Results demonstrate that the sensitivity and precision of the developed method is comparable with most other analytical methods and is prominently rapid than them. We believe that the work provides a novel and effective approach and is beneficial for monitoring and reducing the risk of AA contaminations during the process of fruit juice production.
研究领域:
食品科学
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分子生物学
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高影响因子
JOURNAL OF FOOD COMPOSITION AND ANALYSIS
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2021
·
IF 4.556
Yue-Hong Pang,Yi-Ying Wang,Meng-Meng Sun,Xiao-Fang Shen
The cauliflower mosaicvirus 35S (CaMV35S), as the most common promoter, has a usage rate of 70 % in transgenic crops, especially in dicots. Herein, a visual detection method of rolling circle amplification (RCA) combined with DNAzyme technology was developed for the determination of CaMV35S gene. A template sequence of the G-quadruple gene was designed for combining with the target gene CaMV35S. When the target gene existed, the RCA reaction occurred through the action of the enzyme, and the product combined with hemin to generate DNAzyme, which catalyzed the color reaction of 2′-hydrazine-bis-3-ethylbenzothiazolin-6-sulfonic acid (ABTS). Under the optimal experimental conditions, the linear range is 1 × 10 −13 ∼1 × 10 -8 mol/L, and the detection limit is 3 × 10 -14 mol/L (S/N = 3). The method has been successfully applied to standard samples and practical samples (soybean and tofu). For verifying the feasibility and reliability, real-time fluorescent quantitative PCR method was used to detect the target gene CaMV35S, and the results were in good agreement with those of our method, indicating that our developed method can be used for the determination of actual samples.
研究领域:
分子生物学
·
基因工程
·
农业科学
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高影响因子
JOURNAL OF APPLIED ELECTROCHEMISTRY
·
2022
·Vol. 52
·期 5
·885-893
·
IF 2.925
Zhao Fei,Xue Wenwen,Zhang Hongfang
Abnormal expression and functioning of microRNAs in the immune system are frequently linked to various cancers. Our previous work demonstrated a universal, label-free, and reliable microRNA detection platform based on the nanochannels-based recognition interface and electrode-based sensing interface. Herein, the strategy of G-quadruplex DNAzyme-catalyzed deposition was introduced into the nanochannels to improve the detection sensitivity of the platform. G-quadruplex DNAzyme could catalyze the oxidation of 4-chloro-1-naphthol to produce insoluble precipitates into the nanochannels which amplified the effect of the hybridization reaction between the capture probe and the target to the mass transport of the redox probe through the recognition interface. Then, a porous carbon nanofibers-modified electrode was applied to further amplify the monitoring sensitivity of the sensing interface to the concentration change of the redox probe methylene blue. Thereby, sensitive electrochemical detection of microRNA-21 was acquired. The biosensing platform exhibited a broad linear range of 100 aM to 1 nM for microRNA-21, with a low detection limit of 40 aM. What is more, the selective and reproducible platform provided an alternative technique for the PCR-free determination of microRNA in serum and tumor cells. Graphical abstract
研究领域:
生物化学
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分子生物学
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癌症研究
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电化学
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高影响因子
Frontiers in Chemistry
·
2022
·
IF 5.545
Ding Wen,Miao Qingzhen,Bao Xiuli,Wang Shiyuan,Lu Jing,Lyu Mingsheng,Wang Shujun
Furunculosis, which is caused by Aeromonas salmonicida, can induce septicemia, leading to the rapid death of fishes belonging to Salmonidae, Cyprinidae, and Fuscheridae, and lamprey. Targeting A. salmonicida, five DNAzyme sequences with the highest enrichment rates were selected through the Systematic Evolution of Ligands by Exponential Enrichment (SELEX). The enrichment rates were 34.78, 23.60, 8.91, 2.89, and 2.34%, respectively. The DNAzyme with the highest activity, named D-AS-2, showed specificity and sensitivity. D-AS-2 was combined with carboxyl-functionalized graphene to construct a biosensor, which showed good fluorescence response to scabies lesion samples. The diagnostic procedure was completed in <2 min and can be used for the on-site diagnosis of fish diseases. A low-cost, rapid, simple, and highly specific biosensor for the diagnosis of furunculosis was established based on DNAzyme and carboxyl-functionalized graphene.
研究领域:
水产养殖
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分子生物学
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生物传感器
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PubMed
高影响因子
Sensors and Actuators Reports
·
2025
·
IF 7.6
Haoyue Ma,Ying Liang,Jiangnan Wang,Wanjuan Lin,Yang Pan,Peng Cao,Yue Zhang
The development of colorimetric biosensors offers a promising tool for on-site visual analysis of medicinal plants and related products in market regulation. However, compared to fluorescent or electrochemical sensing strategies, colorimetric sensors often suffer from weak signals and limited sensitivity, making it challenging to detect low-abundance RNA targets in commercial samples. In response to this issue, here we design a triple signal amplification strategy to achieve ultrasensitive colorimetric signal output for visible microRNA (miRNA) detection in medicinal plants and products, and the smartphone-based imaging enabled portable quantitative analysis for on-site market regulation. The specifically expressed miRNA in ginseng is chosen as the target strand to initiate the first amplification (hybridization chain reaction) for DNAzyme activation, catalyzing the successive cleavage of substrate strands for the liberation of trigger strands that drive rolling circle amplification. This process produces a high abundance of repetitive G-quadruplex motifs, which function as catalytic scaffolds to accelerate a colorimetric reaction, ultimately enabling sensitive and reliable detection by the naked eye. Combining three amplification approaches into a unified cascade, this strategy generates distinguishable colorimetric signals with naked eyes, and the quantitative analysis could be achieved using a smartphone software, offering a wide dynamic range (1-10 5 pM) and a low detection limit of 651 fM. Successful application in medicinal plant ginseng and related products demonstrates the practicality for real sample analysis. The modular amplification design holds broad potential for detecting other molecular targets, underscoring its promise for portable biosensing and on-site regulatory applications.
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高影响因子
BIOSENSORS & BIOELECTRONICS
·
2019
·
IF 9.518
Weihua Zhao,Mingbin Liu,Hongbo Li,Suqin Wang,Shasha Tang,Rong-Mei Kong,Ruqin Yu
Accurate and sensitive detection of the Pax-5a gene is of great importance in the early diagnosis and prognosis of acute leukaemia. Herein, a label-free electrochemical sensing system was proposed for the detection of the acute leukaemia Pax-5a gene based on enzyme-assisted signal amplification to generate abundant G-quadruplex/hemin DNAzyme. The presence of Pax-5a can open the hairpin probe (HP), which acts as a template. Under the action of the restriction enzymes Nt.BbvCI and Klenow fragment polymerase , the target gene Pax-5a is cycled to open the HP; On the other hand, a large number of G-quadruplex sequences are produced. The resulting G-quadruplex sequence is capable of forming the G-quadruplex/hemin complex on the surface of the electrode in the presence of hemin . The ultrasensitive label-free electrochemical detection of Pax-5a can be realized via the G-quadruplex/hemin complex-catalysed reduction of H 2 O 2 , and the detection limit was estimated to be as low as 4.6 fM. In addition, the biosensor has good specificity and stability, and also has excellent detection capabilities in a complex substrate environment. Therefore, the sensor shows great potential in bioanalysis and clinical diagnosis.
研究领域:
生物医学
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分子诊断
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电化学传感器
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高影响因子
ANALYTICA CHIMICA ACTA
·
2025
·
IF 6
Lu Lin,Mengmeng Lv,Jiangtao Ren,Jin Wang,Erkang Wang
Background G-quadruplex DNAzymes, known for their peroxidase-like activity and ease of rational design, have emerged as promising tools in biosensing applications. Despite the fundamental importance of structural topology, investigations into how the split mode of intramolecular split G-quadruplex (IntrSG)-based DNAzymes modulates their catalytic properties, as well as their implications for the development of DNAzyme-based probes, remain scarce. In this work, eleven IntrSG strands of different split modes were engineered with a classical parallel G-quadruplex sequence ( PW17 ). The catalytic activity of IntrSG-hemin mixtures was systematically investigated via various techniques. Results Most IntrSG strands incubated with hemin exhibited apparent catalytic activity, which was influenced by split modes and hybridization of inserted spacer. Interestingly, the mixture of hemin and IntrSG of split mode 5:7 (IntrSG(5:7)) exhibited drastic catalytic activity response after hybridization between its spacer and target, and the underlying mechanism was disclosed, which was hybridization-facilitated G-quadruplex assembling and catalytic enhancement. The IntrSG(5:7)-hemin system can be utilized as an effective “signal-on” signal transducer, owing to its simpleness and universality. Subsequently, influence of sequence context changes on the catalytic response was explored by extending and truncating the target sequence. Ignorable effect of protruding DNA was validated, and robust hybridization was critical for catalytic responses, which were unrelated to hybridization location. Finally, an IntrSG(5:7)-based catalytic probe was employed for quantitative analysis of miRNA ( Let-7a ), and favorable performance was achieved. Significance Taken together, a mechanistic framework for topological control of G-quadruplex DNAzyme catalysis was established, highlighting the potential of IntrSG as a versatile platform for biosensing and molecular diagnostics.
研究领域:
生物化学
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分子诊断学
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生物传感
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核酸工程
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DNA纳米技术
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高影响因子
Journal of Pharmaceutical Analysis
·
2025
·
IF 8.9
Zhe Ni,Xingshi Yuan,Zhengliang Luo,Xiaoqi Zhang,Min Chen,Lee Jia,Jie Wang,Xifu Shang
Sensitive detection of microRNA-204 (miR-204) is critical for the early diagnosis and management of osteoarthritis (OA). This work presents a novel surface-enhanced Raman scattering (SERS) biosensor for the ultrasensitive and specific detection of OA-associated miR-204. The platform integrates a self-amplifying nucleic acid circuit with DNAzyme-catalyzed etching of a plasmonic nanoprobe. At its core is a single, rationally designed overhang-containing hairpin probe (O-HP) that functions as both the recognition element and amplification initiator. Upon binding to miR-204, the O-HP triggers polymerase-mediated extension, generating G-quadruplex structures. These structures bind hemin to form DNAzymes that catalyze the localized production of reactive oxygen species (ROS), which subsequently etch the silver shell of the AuNS/Ag@4-ATP SERS nanoprobe. This etching causes the desorption of Raman reporters and a quantifiable 'signal-off' response. This biosensor achieves a remarkably low detection limit of 8.13 fM with a broad dynamic range from 10 fM to 150 nM, and exhibits high specificity, capable of discriminating single-nucleotide variants. Furthermore, it successfully quantified miR-204 in clinical cartilage samples, showing a strong correlation with real-time quantitative polymerase chain reaction results. The modular design of the O-HP also facilitated the adaptation of the platform for detecting miR-21, demonstrating its generalizability. This work provides a robust and versatile biosensing strategy with significant potential for clinical miRNA diagnostics.
研究领域:
生物传感器
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分子诊断
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纳米生物技术
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微小RNA检测
·
即时检测
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高影响因子
TALANTA
·
2026
·
IF 6.1
Tai Ye,Shuying Yue,Long Bai,Haohao Chen,Chenyang Li,Rehemutula Wubuli,Kehua Dang,Min Yuan,Hui Cao,Ziqing Ye,Fei Xu
Co-immobilization of horseradish peroxidase (HRP) with G-quadruplex/hemin complex often suffer from low efficiency, intricate procedures, and reduced catalytic performance. To address these challenges, we developed a biomineralization strategy that integrates HRP and G-quadruplex/hemin into the single nanosheet structure. By engineering a split aptamer sequence into a G-rich domain, the nanosheets synergistically combine enhanced catalytic activity with better target-specific molecular recognition. Compared with either free HRP or it biomineralized counterpart alone, the prepared nanosheets exhibited markedly improved peroxidase-like activity. In the presence of enrofloxacin, specific aptamer-target binding facilitated the formation of a ternary complex on the electrode surface, enabling the catalysis of H 2 O 2 -mediated oxidation of TMB. Under the optimized conditions, the electrochemical aptasensor displayed a wide linear detection range of 1 nM–10 μM with a low detection limit of 0.53 nM. Beyond high selectivity, the sensor demonstrated excellent practical applicability in spiked milk and egg white samples. The generality of the developed platform was further confirmed by its successful extension to other antibiotics detection through replacing the split-aptamer pairs.
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FOOD CONTROL
·
2024
·
IF 5.6
Jiong Liang,Yunzhe Zhang,Zikun Li,Xin Lu,Congyan Qi,Qian Yang,Wei Zhang
Aflatoxin B1 is a potent carcinogen widely found in agricultural products and feed. Therefore, investigating the development of convenient, sensitive, and precise techniques for detecting AFB1 is critical. This work proposes a ratiometric electrochemical aptasensor based on DNAzyme-driven tripedal DNA walker for enzyme-free AFB1 detection. The design of multiple legs reduces the moving time on the electrode surface and enhances the value of signal response. In this approach, the tagging of ferrocene and methylene blue as signal switch molecules diminishes the impact of interferences and significantly enhances the reproducibility (RSD: 2.825%). Meanwhile, the tripedal DNA walker and DNAzyme dual signal amplification enabled the sensor to detect AFB1 traces in cereals. We achieved trace detection of AFB1 with a detection limit of 61 fg/mL and a wide linear range from 0.0001 to 10 ng/mL. Furthermore, the aptasensor success rate was comparable to that of HPLC-PMP. The results demonstrate enormous potential for detection. Furthermore, the method can detect various targets by altering the target aptamer , displaying attractive scalability. This suggests its tremendous prospective in detecting mycotoxins in food in the future.
研究领域:
食品安全检测
·
生物传感技术
·
分子诊断
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DOI
高影响因子
TALANTA
·
2024
·
IF 5.6
Na Li,Xian Li,Xiaoe Ming,Jingyuan Chen,Yeyi Chen,Lifen Zhou,Ruirui Yao,Yuqi Yao
Deferasirox (DEF) is an important iron chelator for treatment of iron overload–related diseases. Monitoring DEF concentration in human serum will provide some valuable information for clinical diagnosis and therapy of such diseases. In this study, we developed a peroxidase-mimicking colorimetric sensor for the detection of DEF by simple assembly of a telomeric dimeric G-quadruplex DNAzyme with Cu 2+ . The DNAzyme–catalyzed oxidation of 3,3′,5,5′-tetramethylbenzidine in the presence of H 2 O 2 can generate a quantitative colorimetric signal, and the color change can be discerned by the naked eye. Compared with the reaction rate of the monomeric G-quadruplex–Cu 2+ DNAzyme, the reaction rate of the dimeric G-quadruplex–Cu 2+ (G2–Cu 2+ ) DNAzyme is significantly accelerated, and the reaction rate gradually increases and then reaches a plateau with increasing number of TTA spacers. Herein, the G2–Cu 2+ DNAzyme is chosen for the highly sensitive detection of DEF based on the DEF–Cu 2+ complex–induced inhibition of its peroxidase-mimicking activities. The limit of detection (LOD) of DEF is achieved as low as 0.03 μM, and the linear range is from 0.05 to 1.2 μM. The proposed strategy exhibits excellent selectivity in the presence of potential interferents, such as metal ions and small molecules. Importantly, the G2–Cu 2+ DNAzyme is further expanded to detect DEF in dispersible tablets and human serum samples. Overall, this G2–Cu 2+ DNAzyme provides a simple, low-cost, and rapid platform for DEF detection. This novel strategy is the first example of DEF analysis by utilizing signal amplification technology based on the G-quadruplex DNAzyme and holds great potential for DEF quality control and therapeutic drug monitoring.
研究领域:
生物化学
·
医学诊断
·
药物分析
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PubMed
高影响因子
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