BIOSENSORS & BIOELECTRONICS
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2025
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IF 10.5
Wen Tian,Tao Liu,Xiaoying Niu,Po Li,Zhehuang Li,Songcheng Yu,Peng Zhang
DNA methylation serves as a critical epigenetic biomarker for early disease screening and clinical diagnostics, exemplified by the FDA-approved detection of methylated Septin9 gene in peripheral blood for colorectal cancer (CRC) screening. Conventional CRISPR/Cas12a-based detection systems rely on fluorophore-quencher-labeled ssDNA probes to transduce trans-cleavage activity into fluorescence signal, which yet suffers from limitations including complex probe synthesis, background noise induced by incomplete quenching and high cost. In this study, we developed a cascaded and label-free CRISPR/Cas12a system for Septin9 methylation detection using DNAzyme and split DAP-10. The method employed methylation-sensitive Aci I digestion to selectively hydrolyze non-methylated DNA, leaving methylated Septin9 DNA fragment intact to activate CRISPR/Cas12a system. Activated Cas12a triggered trans-cleavage of DNAzyme, a bifunctional molecule that acted as both trans-cleavage substrate of Cas12a and hydrolysis regulator of linker DNA that facilitated split DAP-10 reassembly to light-up Auramine O (AO) fluorescence proportionally to methylation levels. The assay achieved a linear range from 2 to 200 nM with a detection limit of 1.74 nM and robust performance in accuracy, precision, and selectivity. The proposed method was successfully applied in methylated Septin9 detection in cell lines, clinical tissues and blood samples, which indicated application potential for CRC screening.
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SPECTROCHIMICA ACTA PART A-MOLECULAR AND BIOMOLECULAR SPECTROSCOPY
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2025
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IF 4.6
Lingchen Wang,Yue Huang,Linlin Sun,Xiaojuan Gong,Chuan Dong,Shaomin Shuang
Herein, a novel rapid, specific, and sensitive probe for adenosine triphosphate (ATP) detection is developed. We have pioneered the integration of split ATP aptamers with aggregation-induced emission (AIE) via chemical binding to obtain a novel ATP detection probe (TPE probe). Upon the binding of the ATP split aptamer to the target, the intermolecular distance of the AIE molecules decreases, restricting their spatial rotation, which leads to an enhancement of the fluorescence signal, thereby enabling the detection of ATP. This unique design provides the probe with both target-specific recognition capability of the aptamer and sensitive signal-reporting function. Under the optimized experimental conditions, the probe exhibits a linearity range from 1-1000 nM with limit of detection 0.14 nM, displaying a comparative advantage over previously reported methods in terms of sensitivity and selectivity. Additionally, this method demonstrated satisfactory performance for ATP detection in beer samples, with recoveries between 97.5 % to 104.6 %. More importantly, the current research is expected to provide an effective monitoring method for beer quality control.
研究领域:
生物化学
·
分析化学
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JOURNAL OF HAZARDOUS MATERIALS
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2022
·
IF 14.224
Shuo Qi,Xiaoze Dong,Yuhan Sun,Yin Zhang,Nuo Duan,Zhouping Wang
DNA walker machines, as one of the dynamic DNA nanodevices, have attracted extensive interest in the field of analysis due to their inherent superiority. Herein, we reported a split aptamer remodeling-initiated target-self-service 3D-DNA walker for ultrasensitive, specific, and high-signal-background ratio determination of 17β-estradiol (E2) in food samples. Two split probes (STWS-a and STWS-b) were rationally designed that can undergo structural reassembled to serve as walking strands (STWS) under the induction of the target. Meanwhile, an intact E6-DNAzyme region was formed and activated at the tail of STWS. The activated E6-DNAzyme then continuously drives the 3D-DNA walker for signal amplification and specific detection of E2. Under optimal conditions, the proposed DNA walker-based biosensor exhibited excellent linearity in the range of 1 pM to 50 nM with a low limit of detection (LOD) of 0.28 pM, and good precision (2.7%) for 11 replicate determinations of 1 nM of E2. Furthermore, the developed DNA walker-based biosensor achieved excellent sensitive analysis of E2 in the complex food matrix with recoveries of 95.6–106.5%. This newly proposed split aptamer-based strategy has the advantages of ultrasensitive, high signal-to-background ratio, and high stability. Noteworthy, the successful operation of the DNA walker initiated by the split aptamer expands the principles of DNA walker design and provides a universal signal amplification platform for trace analysis.
研究领域:
分析化学
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纳米技术
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生物传感器
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分子生物学
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NEW JOURNAL OF CHEMISTRY
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2025
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IF 2.5
Mei Li,Shuanglong Ying,Qian Song,Qingyou Cai
The extensively used tobramycin (TOB) impacts human health and ecological environment. We proposed a smartphone-assisted RGB-dependent method for colorimetrically detecting TOB using dual split aptamer-initiated target hybridization chain reaction (HCR). Four DNA sequences: Sp-a, Sp-b, hairpin H1, and H2 were designed. Sp-a and Sp-b contained one split aptamer sequence and one split trigger sequence, respectively, which specifically bound to target TOB, forming a ternary complex, bringing close the two split triggers. This initiated a trigger-H1 reaction, opening H1, hybridizing with H2, and opening H2. This alternating H1/H2 opening initiated HCR amplification, forming a long double-stranded DNA (dsDNA) structure not binding to gold nanoparticles (AuNPs), causing the unprotected AuNPs and color change. As TOB concentration increased, the system's color shifted from wine to blue, sensitively detecting TOB via colorimetry. Its detection linear ranges for UV-Vis spectrometer and smartphone-based sensing platform were 10 pg/mL-150 ng/mL and 10 pg/mL-150 ng/mL, with LODs being 2.49 pg/mL and 18.52 pg/mL. This high-specificity, high-practicability method showed good recovery rates in actual milk and lake water samples, providing a new on-site and visual TOB detection approach.
研究领域:
分析化学
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生物传感器
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环境监测
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纳米技术
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食品安全
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SENSORS AND ACTUATORS B-CHEMICAL
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2025
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IF 7.7
Huihong Yan,Xuan Luo,Chenglong Bao,Yongsheng Yu,Weiwei Yang
Quinine, a cinchona alkaloid, is utilized as an antimalarial agent or a preservative in aquatic products. Owing to its resistance to biodegradation, quinine readily persists in human body fluids and environmental water systems, posing risks of harmful side effects to human health. Here, we developed a highly sensitive and specific detection strategy for quinine based on a small molecule-templated split aptazyme click ligation reaction (SMT-SpAzyme-CLR). This strategy constructs a highly sensitive fluorescent biosensor by integrating a quinine-specific split aptamer with a Mg 2 + -dependent DNAzyme catalytic core, which undergoes quinine-induced click ligation and activates enzymatic cleavage activity. The recycling of the ligated SpAzyme facilitates a one-to-many signal amplification cascade, substantially enhancing the detection sensitivity. The proposed sensor exhibits a wide linear response range from 0.02 to 50 μM and achieves an impressively low detection limit of 5 nM, outperforming most existing aptasensors. It was successfully applied to the determination of quinine in tonic water samples, with recoveries ranging from 96.9 % to 104.9 %. Furthermore, a lateral flow immunoassay strip was implemented for visual semi-quantitative detection, demonstrating high specificity and strong anti-interference capability even in complex matrices. This work provides a sensing platform that combines the strengths of highly sensitive laboratory quantification and rapid on-site screening for quinine.
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JOURNAL OF MOLECULAR RECOGNITION
·
2019
·Vol. 33
·期 5
·e2829
·
IF 1.919
Shaoyuan Li,Yan Zheng,Yaqin Liu,Xiuhua Geng,Xiaofeng Liu,Liyuan Zou,Qing Wang,Xiaohai Yang,Kemin Wang
Understanding the binding of split aptamer/its target could become a breakthrough in the application of split aptamer. Herein, vascular endothelial growth factor (VEGF), a major biomarker of human diseases, was used as a model, and its interaction with split aptamer was explored with single molecule force spectroscopy (SMFS). SMFS demonstrated that the interaction force of split aptamer/VEGF 165 was 169.44 ± 6.59 pN at the loading rate of 35.2 nN/s, and the binding probability of split aptamer/VEGF 165 was dependent on the concentration of VEGF 165 . On the basis of dynamic force spectroscopy results, one activation barrier in the dissociation process of split aptamer/VEGF 165 complexes was revealed, which was similar to that of the intact aptamer/VEGF 165 . Besides, the dissociation rate constant ( k off ) of split aptamer/VEGF 165 was close to that of intact aptamer/VEGF 165 , and the interaction force of split aptamer/VEGF 165 was higher than the force of intact aptamer/VEGF 165 . It indicated that split aptamer also possessed high affinity with VEGF 165 . The work can provide a new method for exploring the interaction of split aptamer/its targets at single-molecule level.
研究领域:
生物物理学
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分子生物学
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医学诊断
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MICROCHEMICAL JOURNAL
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2022
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IF 5.304
Wenjing Wang,Fei Zhai,Fupei Xu,Min Jia
Novel enzyme-free amplified dual-terminal labeled split aptamer (SPA) probes were developed to achieve one-step rapid and sensitive detection of bisphenol A (BPA). By rational split, the original aptamer was divided into two segments named SPA-a and SPA-b to achieve target-driven self-assembly and form BPA/SPA complex. To achieve enzyme-free signal amplification and improved sensitivity, SPA probes were dual-terminal labeled with 6-carboxyfluorescein (FAM) fluorophores and black hole quencher 2 (BHQ 2 ). Two dual-terminal labeled SPA probes were induced to assemble a ternary complex with BPA to produce fluorescence resonance energy transfer effect, generating the decreased fluorescence intensity. The detection limit of dual-terminal labeled SPA probes was 0.89 ng/mL, which was 24-fold lower than that of single-terminal modification. Meanwhile, these SPA probes showed a rapid response to BPA, which could be completed in 5 min for one-step detection without complex cleaning and separation progress. The recovery rates in real samples were 94.17% to 107.28%. In brief, the dual-terminal labeled SPA probes showed substantial potential for onsite detection of BPA.
研究领域:
分析化学
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环境科学
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材料科学
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FOOD CHEMISTRY
·
2024
·
IF 8.5
Xin Zheng,Zunqi Hu,Shunxiang Gao,Zhen Li,Jia Chen,Guanyi Zhang,Nana Kong,Jianguo Sun,Weiwei Liu
An aptamer targeting gliotoxin (GTX) was optimized to increase the binding affinity by approximately 20 times and achieve higher structural stability and targeting specificity. Molecular dynamics simulations were used to explore the molecular mechanism and key action sites underlying the recognition of GTX by the optimized aptamer . Subsequently, the optimized aptamer was split into two fragments and a convenient and rapid one-pot assay for GTX detection was successfully established using a target-driven split aptamer recognition and assembly strategy. The method exhibited a good linear range of 0.128 nM to 2 μM, a low detection limit of 0.07 nM, and excellent selectivity for GTX. Furthermore, the method had good accuracy and stability in real sample analysis. Therefore, the developed one-pot method provides a reliable, convenient, and cost-effective approach for the widespread application of GTX detection.
研究领域:
生物化学
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分析化学
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毒理学
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TALANTA
·
2026
·
IF 6.7
Chenglong Chen,Qiaoxia Zhang,Xiajuan Wang,Ran Miao,Tiesheng Shi,Qingyun Liu
To resolve the conflict between multidimensional interferences derived from complex matrices and the stringent requirements for trace-level detection sensitivity, a novel electrochemical aptasensor was constructed for highly sensitive kanamycin (KANA) detection based on a split aptamer engineering strategy. Ag–Ti 3 C 2 T x MXene, synthesized via in-situ reduction of silver nanoparticles onto the surface of two-dimensional MXene nanosheets, is employed for electrode decoration. This composite possesses abundant binding sites for aptamer 1 (Apt1) immobilization and exhibits outstanding catalytic performance in the reduction of H 2 O 2 . Signal acquisitions were performed under reductive potential conditions, endowing the sensor with superior anti-interference capability. Upon KANA addition, aptamer 2 (Apt2) can assemble into a ternary structure with the target and Apt1, leading to the displacement of cDNA and triggering a signal-down response. Under optimized conditions, the fabricated aptasensor presents a broad linear range of 1∼20000 nM with a limit of detection (LOD) of 0.12 nM, and exhibits excellent selectivity, reproducibility, and strong stability. In addition, the aptasensor demonstrates satisfactory recovery rates in real sample analysis, indicating its great potential for practical applications. The combination of Ag–Ti 3 C 2 T x MXene nanocomposite and split aptamer engineering technology provides a feasible strategy for highly sensitive detections of trace-level antibiotic residues in complex food matrices.
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TALANTA
·
2026
·
IF 6.7
Ziyu Jiang,Danting Yang,Meng Luo,Hailiang Qin,Yanru Fan,Shuxia Xu
Residual kanamycin (Kana) in water bodies threatens ecological safety and human health, driving demand for straightforward and sensitive detection tools. In this work, we present a label-free electrochemical sandwich aptasensor that achieves ultrasensitive Kana detection by synergizing split-aptamer recognition with signal amplification through flower-like gold nanostructures (AuNFs). The split aptamer fragments (SPA1/SPA2) remain separate in the absence of target, minimizing background from nonspecific folding. Upon Kana introduction, a SPA1–Kana–SPA2 ternary complex assembles on the AuNF-modified ITO electrode, providing a scaffold for intercalation of the redox indicator ferrocene–naphthalimide (FND). The consequent accumulation of FND results in a markedly enhanced differential pulse voltammetry response. The three-dimensional AuNFs offer a large electroactive surface for high-density probe immobilization and efficient electron transfer. The sensor exhibits a linear response for Kana from 5 nM to 2 μM with a detection limit of 0.2 nM (3 σ / k ). It also shows excellent selectivity over analogous aminoglycosides, good reproducibility, and acceptable storage stability. The aptasensor was successfully applied to the analysis of real water samples, yielding recoveries of 86.4–116.8%, consistent with HPLC-MS measurements. This work provides a sensitive, selective, and reliable platform for on-site monitoring of Kana in water.
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RSC Advances
·
2015
·Vol. 5
·期 92
·75746-75752
·
IF 3.84
Siqi Zhang,Kun Wang,Jiali Li,Zhenyu Li,Ting Sun
In this study, a highly efficient colorimetric assay for adenosine triphosphate (ATP) was constructed based on a split aptamer target binding strategy and grapheme oxide–platinum/gold nanoparticle platform to increase catalytic properties. Platinum/gold nanoparticles were easily prepared by reducing the mixture solution of K2PtCl6 and HAuCl4 using sodium citrate in a one-pot synthesis. Next, graphene oxide/platinum/gold nanoparticle hybrids (GO/PDDA/PtAuNPs) were fabricated using a self-assembly procedure. Ultrasensitive and enzyme-free detection of ATP in fetal bovine serum was achieved using split aptamer-modified magnetic beads as a separating unit and split aptamer-modified GO/PDDA/PtAuNPs as a catalyst-detecting platform. The specific binding of ATP and its aptamer linked the split aptamer-modified GO/PDDA/PtAuNPs and magnetic beads together. Using magnetic separation, achromatous tetramethylbenzidine was catalyzed into a colored product by separated nanocomposites, which enabled rapid detection of ATP. The color change caused by the low concentration ATP (50 nM) was clearly distinguished by direct observation. The detection limit of the colorimetric assay for ATP was 0.2 nM, which is close to the lowest detection limit for colorimetric detection of ATP. This assay showed excellent selectivity against guanosine triphosphate, uridine triphosphate, and cytosine triphosphate and can be applied in real samples.
研究领域:
分析化学
·
纳米技术
·
生物化学
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