MICROCHIMICA ACTA
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2024
·Vol. 191
·期 7
·1-11
·
IF 5.3
Liang Pengda,Lv Bei,Chen Ke,Li Dawei
The combination of CRISPR/Cas12a and functional DNA provides the possibility of constructing biosensors for detecting non-nucleic-acid targets. In the current study, the duplex protospacer adjacent motif (PAM) in the activator of CRISPR/Cas12a was used as a molecular switch, and a sensitive adenosine triphosphate (ATP) detection biosensor was constructed using an allosteric probe-conjugated PAM site formation in hybridization chain reaction (HCR) integrated with the CRISPR/Cas12a system (APF-CRISPR). In the absence of ATP, an aptamer-containing probe (AP) is in a stem-loop structure, which blocks the initiation of HCR. In the presence of ATP, the structure of AP is changed upon ATP binding, resulting in the release of the HCR trigger strand and the production of long duplex DNA with many PAM sites. Since the presence of a duplex PAM site is crucial for triggering the cleavage activity of CRISPR/Cas12a, the ATP-dependent formation of the PAM site in HCR products can initiate the FQ-reporter cleavage, allowing ATP quantification by measuring the fluorescent signals. By optimizing the sequence elements and detection conditions, the aptasensor demonstrated superior detection performance. The limit of detection (LOD) of the assay was estimated to be 1.16 nM, where the standard deviation of the blank was calculated based on six repeated measurements. The dynamic range of the detection was 25–750 nM, and the whole workflow of the assay was approximately 60 min. In addition, the reliability and practicability of the aptasensor were validated by comparing it with a commercially available chemiluminescence kit for ATP detection in serum. Due to its high sensitivity, specificity, and reliable performance, the APF-CRISPR holds great potential in bioanalytical studies for ATP detection. In addition, we have provided a proof-of-principle for constructing a CRISPR/Cas12a-based aptasensor, in which the PAM is utilized to regulate Cas12a cleavage activity. Graphical Abstract
研究领域:
生物化学
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分子生物学
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分析化学
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PubMed
高影响因子
Journal of Analysis and Testing
·
2023
·
IF 4.7
Gong Xue,Wu Jiangling,Zhang Jiajia,Jiang Zhongwei,Wang Yi,Zhang Pu
Heart failure (HF) is a clinical hallmark of cardiovascular syndrome leading to substantial morbidity, repeated hospitalization, and enormous healthcare expenditure. As elderly populations continue to increase, biomarker-guided diagnosis and treatment for age-related cardiac decline have become essential. Accumulating evidence demonstrates that N-terminal proBNP (NT-proBNP) can provide a unique window into the diagnosis and risk stratification with HF. Herein, a simple yet robust aptasensor is developed for on-site recognizing the NT-proBNP by its targeting aptamer, thus achieving the accurate diagnosis of HF. This aptasensing system is prepared by absorbing the fluorophore-labeled aptamer strand onto the graphene oxide (GO), leading to efficient quenching without possible off-site signal leakage. The aptamer strand can specifically identify target NT-proBNP molecules via a versatile conformational transformation, resulting in the desorption of the aptamer-NT-proBNP complexes from GO and re-generation fluorescence signal, thus allowing sensitive detection of NT-proBNP in 37 clinical blood samples. Taking advantage of the high specificity of aptamer-guaranteed recognitions of NT-proBNP, this aptasensor system readily achieves better diagnostic performance for HF than commercially adopted chemiluminescence immunoassay (Siemens, CLIA) in hospitals in terms of accuracy (89.2% vs 83.8%), specificity (89.5% vs 84.2%), and positive predictive value (88.9% vs 83.3%). This work provides a stable option for the diagnosis and treatment of elderly HF-related diseases in clinics.
研究领域:
医学
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生物技术
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DOI
高影响因子
JOURNAL OF HAZARDOUS MATERIALS
·
2025
·
IF 11.3
Wenjing Wang,Rui Feng,Kaiyue Wei,Jiashuo Xu,Wenhui Dong,Jiao Li,Jianan Sun,Sai Wang,Xiangzhao Mao
Excessive histamine in spoiled seafood poses considerable health hazards to consumers, yet its detection is challenging due to complicated sample preparation and detection methodologies. Herein, an integrated colorimetric platform containing Poly (vinyl alcohol) (PVA)/ hyaluronic acid (HA) microneedle patchs-assisted extraction and aptasensor-based detection was reported. The developed PVA/HA microneedle patches facilitated on-site histamine extraction from seafood through a two-minute press-and-peel procedure. To enhance detection efficacy, strategies for generating high-affinity aptamers with specific terminal-fixed structures and constructing AuNPs@FeP-chitosan oligosaccharide (COS) nanozyme boosting catalytic efficiency were proposed. Utilizing the aptamer HIS3-T2 in conjunction with the nanozyme, a colorimetric aptasensor was developed. Integrated with the patches, the aptasensor achieved high sensitivity and selectivity, detecting histamine within a range of 2~800 nM with a limit of detection (LOD) of 1.89 nM. Validated on real-world salmon and shrimp samples, this integrated system promises rapid and accurate histamine monitoring, offering great reference for similar applications in food quality control.
研究领域:
生物传感技术
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食品安全
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分析化学
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DOI
PubMed
高影响因子
MOLECULES
·
2022
·Vol. 27
·期 6
·1871
·
IF 4.927
Jing Yang,Wei Zhong,Qi Yu,Jin Zou,Yansha Gao,Shuwu Liu,Songbai Zhang,Xiaoqiang Wang,Limin Lu
A simple and label-free electrochemical aptasensor was developed for ultra-sensitive determination of chloramphenicol (CAP) based on a 2D transition of metal carbides (MXene) loaded with gold nanoparticles (AuNPs). The embedded AuNPs not only inhibit the aggregation of MXene sheets, but also improve the quantity of active sites and electronic conductivity. The aptamers (Apts) were able to immobilize on the MXene–AuNP modified electrode surface through Au–S interaction. Upon specifically binding with CAP with high affinity, the CAP–Apt complexes produced low conductivity on the aptasensor surface, leading to a decreased electrochemical signal. The resulting current change was quantitatively correlated with CAP concentration. Under optimized experimental conditions, the constructed aptasensor exhibited a good linear relationship within a wide range of 0.0001–10 nM and with a low detection limit of 0.03 pM for CAP. Moreover, the developed aptasensor has been applied to the determination of CAP concentration in honey samples with satisfactory results.Keywords:MXene;gold nanoparticle;chloramphenicol;electrochemical;aptasensor
研究领域:
分析化学
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纳米技术
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食品安全
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DOI
PubMed
高影响因子
ANALYTICAL CHEMISTRY
·
2020
·
IF 6.785
Weiwei Qiu,Feng Gao,Natsumi Yano,Yusuke Kataoka,Makoto Handa,Weiqiang Yang,Hidekazu Tanaka,Qingxiang Wang
Electrochemical aptasensors involved in chemical labeling are often single-use and sensitivity-limited because the probes are commonly single-point labeled and irreversible. In this work, the specific coordination between Zr4+ and phosphate group (−PO43–) was employed to construct a new aptasensor that is highly sensitive and reusable, using Ochratoxin A (OTA) as the test model. The OTA binding aptamer (OBA) was hybridized with the thiolated supporting sequence (TSS) immobilized on the surface of a gold electrode. The UiO-66 with a formula of [Zr6O4(OH)4(BDC)6], one of the class of Zr metal–organic frameworks (MOFs), was then particularly grafted on the terminal of OBA through the specific coordination between Zr4+ and 5′-PO43–, i.e., the Zr–O–P coordination bond. Similarly, as much as the 5′-PO43– and 3′-methylene blue dual-labeled sequences (DLS) were further assembled on UiO-66 due to the large surface area of MOF and rich active sites of Zr4+. Owing to the specific coordination for signal amplification, the developed aptasensor shows greatly enhanced sensitivity. A wide detection range from 0.1 fM to 2.0 μM and an ultralow detection limit of 0.079 fM (S/N = 3) for OTA were obtained. Additionally, the TSS can rehybridize with a new OBA to regenerate the aptasensor but without complicated pretreatments, enabling a aptasensor that is readily reusable for OTA detection. The aptasensor was successfully applied for OTA detection in the red wine samples, demonstrating a promising prospect for food safety monitoring.
研究领域:
生物化学
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食品安全检测
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纳米材料科学
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DOI
高影响因子
JOURNAL OF FOOD PROCESS ENGINEERING
·
2024
·Vol. 47
·期 6
·e14654
·
IF 2.7
Chengquan Wang,Chengdong Gu,yanna Rong,Xin Zhao,Lu Qian,Mengting Liu,Xingyi Huang,Jing Qian
In this work, a paper-based electrochemical aptamer sensor was developed for the detection of aflatoxin B1 (AFB1) using a combination of MXene–Ti 3 C 2 T x and nucleic acid aptamers. The prepared single-layer or few-layer MXene suspension is suction-filtered onto MXene paper, which is cut to prepare MXene electrodes. To accomplish AFB1 specific detection, an amino-labeled AFB1 aptamer is mounted on the surface of the carboxy-functionalized MXene electrode. When AFB1 is present, it particularly binds to the aptamer to form a 3D structure, reducing the efficiency of electron transmission on the sensor surface. The difference in impedance signal change at the electrode/electrolyte interface is used to quantify AFB1. The results indicated that the detection range is 0.05–100 ng/mL, the detection limit is 0.04 ng/mL, and the recovery rate of AFB1 in corn samples is 97.8%–111.52% with the optimal detection conditions. The MXene paper-based label-free aptasensor is versatile and can detect different targets by simply swapping out the aptamers of different targets. The sensor also has a wide range of applications in food analysis and environmental testing. Practical applications A paper-based electrochemical aptamer sensor was developed to detect aflatoxin B1 using a combination of MXene–Ti 3 C 2 T x and nucleic acid aptamers.The design is based on the preparation of MXene electrodes by pumping and filtering monolayer or multilayer MXene suspensions onto MXene paper and cutting.The MXene paper-based label-free aptamer sensor was designed to be versatile, allowing the detection of different targets by simply replacing the aptamer with one from a different targets.
研究领域:
食品安全
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环境科学
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生物传感技术
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DOI
高影响因子
ANALYTICAL CHEMISTRY
·
2021
·
IF 6.986
Danli Hu,Haiyan Cui,Xinyang Wang,Fang Luo,Bin Qiu,Weicheng Cai,Hui Huang,Jian Wang,Zhenyu Lin
An Au/GaN photoelectrode was prepared by sputtering 30 nm thick Au film on the surface of n-type gallium nitride (GaN). When the electrode contacts with multilayered molybdenum disulfide (MoS2), photogenerated electrons and photogenerated holes transfer to MoS2 because of the band gap matching of MoS2 and GaN. The presence of Au promotes charge transfer and results in a greater recombination of electrons and holes; by this means, a more significant suppression of photocurrent can be detected. This characteristic has been coupled with the high selectivity of an aptamer and applied to develop a novel photoelectrochemical aptasensor for cancer biomarkers (alpha-fetoprotein (AFP) as a model). The aptamer of AFP was modified on the surface of the Au/GaN photoelectrode by Au–S bonds, which can bind to the target protein with high selectivity. Then, the transfer process of the charge carriers of GaN to MoS2 can be blocked by the target protein so that the suppression of photocurrent is reduced. The difference of the photocurrent in the presence and absence of AFP (ΔI) showed a linear relationship with AFP concentration that ranged from 1.0–150 ng/mL (R2 = 0.9995), and the detection limit was 0.3 ng/mL. The standard addition recovery rates ranged from 85.2 to 91.7%. The method possessed good sensitivity and high selectivity for AFP detection. The developed biosensor can be modified to detect other cancer biomarkers by simply replacing the aptamer used.
研究领域:
生物传感技术
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癌症诊断
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材料科学
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DOI
PubMed
高影响因子
JOURNAL OF FOOD COMPOSITION AND ANALYSIS
·
2023
·
IF 4.3
Baihui Wang,Huan Jiang,Ruyi Tang,Yaoyu Tan,Xuemin Xia,Xia Zhang
The activity of nanozymes is dependent on their surface for catalysis, and bioconjugation or adsorption of DNA aptamers will promote their activity. This paper aims to develop a novel aptamer sensor, for rapid, sensitive and specific detection of histamine. The design of the aptamer biosensor is based on the adsorption of histamine aptamers onto the surface of Au NPs by bioconjugation, which promotes the peroxidase activity of the nanozymes. Since histamine is strongly selective and specific for histamine aptamers, the histamine aptamer can be detached from the surface of Au NPs and bound to histamine, during which the catalytic activity of the nanozymes falls back due to the detachment of the histamine aptamer, which can be quantitative analysis by the TMB-H 2 O 2 system. The decrease in enzyme catalytic activity became larger with increasing histamine concentration, with a good linear correlation. Finally, the histamine aptamer biosensor was constructed, which has the advantages of ultrasensitive, high-accuracy, and wide linear range. The linear range and detection limit are 10 −11 ∼10 −3 M and 1.22 × 10 −12 M, respectively. With reference to this aptamer sensor design, this approach can also be extended to other schemes where the aptamer is combined with Au NPs as a way to detect the corresponding molecules.
研究领域:
生物化学
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分析化学
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纳米技术
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DOI
高影响因子
MOLECULES
·
2023
·Vol. 28
·期 16
·6074
·
IF 4.6
Fang Li,Shuyue Xiong,Pei Zhao,Panpan Dong,Zijian Wu
Quantitative detection of veterinary drug residues in animal-derived food is of great significance. In this work, a simple and label-free electrochemical aptasensor for the highly sensitive detection of chloramphenicol (CAP) in milk was successfully developed based on a new biosensing method, where the single- or few-layer Ti3C2MXene nanosheets functionalized via the specific aptamer by self-assembly were used as electrode modifiers for a glassy carbon electrode (aptamer/Ti3C2MXene/GCE). Differential pulse voltammetry (DPV), electrochemical impedance spectroscopy (EIS), cyclic voltammetry (CV), scanning electron microscopy (SEM), atomic force microscope (AFM), and so on were utilized for electrochemical and morphological characterization. Under the optimized conditions, the constructed aptasensor exhibited excellent performance with a wider linearity to CAP in the range from 10 fM to 1 μM and a low detection limit of 1 fM. Aptamer/Ti3C2MXene/GCE demonstrated remarkable selectivity over other potentially interfering antibiotics, as well as exceptional reproducibility and stability. In addition, the aptasensor was successfully applied to determine CAP in milk with acceptable recovery values of 96.13% to 108.15% and relative standard deviations below 9%. Therefore, the proposed electrochemical aptasensor is an excellent alternative for determining CAP in food samples.Keywords:label-free;electrochemical aptasensor;chloramphenicol;Ti3C2MXene nanosheets
研究领域:
分析化学
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食品安全
·
电化学
·
纳米技术
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DOI
PubMed
高影响因子
FOOD CHEMISTRY
·
2023
·
IF 8.8
Jiaojiao Zhou,Xuqin Lv,Yue Gui,Jiangling He,Fang Xie,Jie Cai
Ochratoxin A (OTA) is a powerful mycotoxin present in a variety of food products, and its detection is important for human health. Here, a fluorescent aptasensor is reported for sensitive OTA determination. Specifically, the surface of bio-inspired passion fruit-like dendritic mesoporous silica nanospheres-enriched quantum dots (MSNQs-apt) was first modified with the OTA aptamer as the recognition unit and fluorescence emitter, while the aptamer-complementary DNA (MNPs-cDNA) was linked with the magnetic nanoparticles (MNPs) as the separation element. In the range of 2.56 pg/mL to 8 ng/mL, the proposed aptasensor exhibited satisfactory linearity and a detection limit of 1.402 pg/mL. The developed aptasensor achieved recoveries of 90.98–103.20% and 94.33–107.57 % in red wine and wheat flour samples, respectively. By simply replacing the aptamer, this aptasensor can be easily extended to detection of other analytes, suggesting its potential as a universal detection platform for mycotoxins in food products.
研究领域:
食品科学
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分析化学
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纳米技术
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毒理学
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DOI
PubMed
高影响因子
MICROCHIMICA ACTA
·
2020
·Vol. 187
·期 4
·1-8
·
IF 6.232
Li Jingxian,Shan Xueling,Jiang Ding,Wang Wenchang,Chen Zhidong
Ultrasensitive and selective determination of bisphenol A (BPA) was achieved by a solid-state electrochemiluminescence (ECL) aptasensor. The Ti-based MOFs (MIL-125, MIL stands for Materials of Institute Lavoisier) have porous structure and large specific surface area. They were used as a carrier for Ru(bpy) 3 2+ by a hydrothermal synthesis method. The Ru(bpy) 3 2+ encapsulated MIL-125 (Ru(bpy) 3 2+ @MIL-125) was dropped on a glass carbon electrode (GCE) and used as working electrode. After that, the thiol-based aptamer of BPA was used to modify the working electrode by Ti–S bond. As a result of the specific binding affinity between aptamer and BPA, the ECL signal was significantly quenched. Thus, a selective electrochemiluminescence (ECL) aptasensor for BPA was developed. The ECL aptasensor based on Ru(bpy) 3 2+ @MIL-125 has a good ECL response toward detecting BPA. Under optimal conditions, the aptasensor can detect BPA in a wide linear range from 1.0 × 10 −12 to 1.0 × 10 −6 M with an excellent determination limit of 6.1 × 10 −13 M (S/ N = 3). The ECL aptasensor can selectively detect BPA in a mixture containing BPA interference compounds.
研究领域:
分析化学
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环境科学
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DOI
PubMed
高影响因子
ANALYTICAL AND BIOANALYTICAL CHEMISTRY
·
2024
·
IF 3.8
Zhang Runze,Wang Yuzhu,Wang Xiaoxu,Ren Honglin,Du Junzheng,Yang Yongjie,Hu Xueyu,Shi Ruoran,Zhang Bo,Li Chengwei,Lu Shiying,Li Yansong,Liu Zengshan,Hu Pan
Listeria monocytogenes ( L. monocytogenes ) is a prevalent food-borne pathogen that can cause listeriosis, which manifests as meningitis and other symptoms, potentially leading to fatal outcomes in severe cases. In this study, we developed an aptasensor utilizing carboxylated magnetic beads and Cas12a to detect L. monocytogenes . In the absence of L. monocytogenes , the aptamer maintains its spatial configuration, keeping the double-stranded DNA attached and preventing the release of a startup template and activation of Cas12a’s trans-cleavage capability. Conversely, in the presence of L. monocytogenes , the aptamer undergoes a conformational change, releasing the double-stranded DNA to serve as a startup template, thereby activating the trans-cleavage capability of Cas12a. Consequently, as the concentration of L. monocytogenes increases, the observable brightness in a blue light gel cutter intensifies, leading to a rise in fluorescence intensity difference compared to the control. This Cas12a aptasensor demonstrates excellent sensitivity towards L. monocytogenes , with a lowest detection limit (LOD) of 57.15 CFU/mL and a linear range of 4×10 2 to 4×10 7 CFU/mL ( R 2 =0.9858). Notably, the proposed Cas12a aptasensor exhibited outstanding selectivity and recovery in beef samples, and could be employed for precise monitoring. This Cas12a aptasensor not only provides a novel fluorescent and visual rapid detection method for L. monocytogenes but also offers simplicity, speed, and stability compared to previous detection methods. Furthermore, it is suitable for on-site detection of beef samples. Graphical Abstract
研究领域:
微生物学
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分子生物学
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食品科学
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PubMed
高影响因子
危险品化学品经营许可证（带存储）