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Boosting Electrochemical Sensing Performances Using Molecularly Imprinted Nanoparticles 利用分子印迹纳米粒子提升电化学传感性能
Pub Date : 2024-07-22 DOI: 10.3390/bios14070358
Francesco Gagliani, Tiziano Di Giulio, Muhammad Ibrar Asif, Cosimino Malitesta, Elisabetta Mazzotta
Nanoparticles of molecularly imprinted polymers (nanoMIPs) combine the excellent recognition ability of imprinted polymers with specific properties related to the nanosize, such as a high surface-to-volume ratio, resulting in highly performing recognition elements with surface-exposed binding sites that promote the interaction with the target and, in turn, binding kinetics. Different synthetic strategies are currently available to produce nanoMIPs, with the possibility to select specific conditions in relation to the nature of monomers/templates and, importantly, to tune the nanoparticle size. The excellent sensing properties, combined with the size, tunability, and flexibility of synthetic protocols applicable to different targets, have enabled the widespread use of nanoMIPs in several applications, including sensors, imaging, and drug delivery. The present review summarizes nanoMIPs applications in sensors, specifically focusing on electrochemical detection, for which nanoMIPs have been mostly applied. After a general survey of the most widely adopted nanoMIP synthetic approaches, the integration of imprinted nanoparticles with electrochemical transducers will be discussed, representing a key step for enabling a reliable and stable sensor response. The mechanisms for electrochemical signal generation will also be compared, followed by an illustration of nanoMIP-based electrochemical sensor employment in several application fields. The high potentialities of nanoMIP-based electrochemical sensors are presented, and possible reasons that still limit their commercialization and issues to be resolved for coupling electrochemical sensing and nanoMIPs in an increasingly widespread daily-use technology are discussed.
分子印迹聚合物纳米粒子(nanoMIPs)结合了印迹聚合物的卓越识别能力和与纳米尺寸有关的特殊性质,如高表面体积比,从而产生了具有表面暴露结合位点的高性能识别元件,促进了与目标的相互作用,进而提高了结合动力学。目前有不同的合成策略来生产纳米 MIPs,可以根据单体/模板的性质选择特定的条件,更重要的是可以调整纳米粒子的大小。优异的传感特性,加上适用于不同目标的尺寸、可调谐性和合成方案的灵活性,使得纳米 MIPs 在传感器、成像和药物输送等多个领域得到广泛应用。本综述总结了纳米 MIPs 在传感器中的应用,尤其侧重于电化学检测,纳米 MIPs 在这方面的应用最为广泛。在对最广泛采用的纳米 MIP 合成方法进行总体调查之后,将讨论印迹纳米粒子与电化学传感器的整合,这是实现可靠、稳定的传感器响应的关键步骤。此外,还将对电化学信号的产生机制进行比较,并说明基于纳米 MIP 的电化学传感器在多个应用领域的应用情况。介绍了基于纳米 MIP 的电化学传感器的巨大潜力,并讨论了仍然限制其商业化的可能原因,以及将电化学传感和纳米 MIPs 结合到日益广泛的日常使用技术中需要解决的问题。
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引用次数: 0
AI-Assisted Detection of Biomarkers by Sensors and Biosensors for Early Diagnosis and Monitoring 人工智能辅助传感器和生物传感器检测生物标记物,用于早期诊断和监测
Pub Date : 2024-07-22 DOI: 10.3390/bios14070356
Tomasz Wasilewski, Wojciech Kamysz, Jacek Gębicki
The steady progress in consumer electronics, together with improvement in microflow techniques, nanotechnology, and data processing, has led to implementation of cost-effective, user-friendly portable devices, which play the role of not only gadgets but also diagnostic tools. Moreover, numerous smart devices monitor patients’ health, and some of them are applied in point-of-care (PoC) tests as a reliable source of evaluation of a patient’s condition. Current diagnostic practices are still based on laboratory tests, preceded by the collection of biological samples, which are then tested in clinical conditions by trained personnel with specialistic equipment. In practice, collecting passive/active physiological and behavioral data from patients in real time and feeding them to artificial intelligence (AI) models can significantly improve the decision process regarding diagnosis and treatment procedures via the omission of conventional sampling and diagnostic procedures while also excluding the role of pathologists. A combination of conventional and novel methods of digital and traditional biomarker detection with portable, autonomous, and miniaturized devices can revolutionize medical diagnostics in the coming years. This article focuses on a comparison of traditional clinical practices with modern diagnostic techniques based on AI and machine learning (ML). The presented technologies will bypass laboratories and start being commercialized, which should lead to improvement or substitution of current diagnostic tools. Their application in PoC settings or as a consumer technology accessible to every patient appears to be a real possibility. Research in this field is expected to intensify in the coming years. Technological advancements in sensors and biosensors are anticipated to enable the continuous real-time analysis of various omics fields, fostering early disease detection and intervention strategies. The integration of AI with digital health platforms would enable predictive analysis and personalized healthcare, emphasizing the importance of interdisciplinary collaboration in related scientific fields.
随着消费类电子产品的稳步发展,以及微流技术、纳米技术和数据处理技术的改进,经济高效、用户友好的便携式设备应运而生,它们不仅是小工具,也是诊断工具。此外,许多智能设备还能监测病人的健康状况,其中一些还被应用于护理点(PoC)测试,作为评估病人病情的可靠来源。目前的诊断方法仍以实验室测试为基础,首先要收集生物样本,然后由训练有素的人员使用专业设备在临床条件下进行测试。实际上,实时收集患者的被动/主动生理和行为数据并将其输入人工智能(AI)模型,可以省略传统的取样和诊断程序,同时也排除了病理学家的作用,从而显著改善诊断和治疗程序的决策过程。将传统和新型的数字和传统生物标志物检测方法与便携式、自主式和微型化设备相结合,可在未来几年内彻底改变医疗诊断方法。本文重点比较了传统临床实践与基于人工智能和机器学习(ML)的现代诊断技术。所介绍的技术将绕过实验室并开始商业化,从而改进或替代当前的诊断工具。将这些技术应用于 PoC 环境或作为消费者技术提供给每一位患者似乎是一种真正的可能。预计未来几年该领域的研究将不断加强。传感器和生物传感器方面的技术进步预计将实现对各种 omics 领域的连续实时分析,促进早期疾病检测和干预策略。人工智能与数字健康平台的整合将实现预测分析和个性化医疗保健,强调了相关科学领域跨学科合作的重要性。
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引用次数: 0
CLICK-FLISA Based on Metal–Organic Frameworks for Simultaneous Detection of Fumonisin B1 (FB1) and Zearalenone (ZEN) in Maize 基于金属有机框架的 CLICK-FLISA 法同时检测玉米中的伏马菌毒素 B1 (FB1) 和玉米赤霉烯酮 (ZEN)
Pub Date : 2024-07-21 DOI: 10.3390/bios14070355
Jingyang Zhang, Banglei Zhu, Xiaoyu Zhang, Yuan Peng, Shuang Li, Dianpeng Han, Shuyue Ren, Kang Qin, Yu Wang, Huanying Zhou, Zhixian Gao
Mycotoxins are secondary products produced primarily by fungi and are pathogens of animals and cereals, not only affecting agriculture and the food industry but also causing great economic losses. The development of rapid and sensitive methods for the detection of mycotoxins in food is of great significance for livelihood issues. This study employed an amino-functionalized zirconium luminescent metal–organic framework (LOF) (i.e., UiO-66-NH2). Click chemistry was utilized to assemble UiO-66-NH2 in a controlled manner, generating LOF assemblies to serve as probes for fluorescence-linked immunoassays. The proposed fluoroimmunoassay method for Zearalenone (ZEN) and Fumonisin B1 (FB1) detection based on the UiO-66-NH2 assembled probe (CLICK-FLISA) afforded a linear response range of 1–20 μmol/L for ZEN, 20 μmol/L for FB1, and a very low detection limit (0.048–0.065 μmol/L for ZEN; 0.048–0.065 μmol/L for FB1). These satisfying results demonstrate promising applications for on-site quick testing in practical sample analysis. Moreover, the amino functionalization may also serve as a modification strategy to design luminescent sensors for other food contaminants.
霉菌毒素是主要由真菌产生的次级产物,是动物和谷物的病原体,不仅影响农业和食品工业,还造成巨大的经济损失。开发快速灵敏的方法来检测食品中的霉菌毒素对解决民生问题具有重要意义。本研究采用了一种氨基功能化锆发光金属有机框架(LOF)(即 UiO-66-NH2)。通过点击化学反应,以可控方式组装 UiO-66-NH2,生成 LOF 组合物,作为荧光连接免疫分析的探针。基于 UiO-66-NH2 组装探针的拟议玉米赤霉烯酮(ZEN)和伏马菌素 B1(FB1)检测荧光免疫分析方法(CLICK-FLISA)的线性响应范围为 1-20 μmol/L(ZEN)和 20 μmol/L(FB1),检测限非常低(ZEN 为 0.048-0.065 μmol/L;FB1 为 0.048-0.065 μmol/L)。这些令人满意的结果表明,在实际样品分析中现场快速检测的应用前景广阔。此外,氨基功能化也可作为一种改性策略,用于设计其他食品污染物的发光传感器。
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引用次数: 0
Allergen Microarrays and New Physical Approaches to More Sensitive and Specific Detection of Allergen-Specific Antibodies 过敏原微阵列和新的物理方法,更灵敏、更特异地检测过敏原特异性抗体
Pub Date : 2024-07-20 DOI: 10.3390/bios14070353
Pavel Sokolov, Irina Evsegneeva, Alexander Karaulov, Alyona Sukhanova, Igor Nabiev
The prevalence of allergic diseases has increased tremendously in recent decades, which can be attributed to growing exposure to environmental triggers, changes in dietary habits, comorbidity, and the increased use of medications. In this context, the multiplexed diagnosis of sensitization to various allergens and the monitoring of the effectiveness of treatments for allergic diseases become particularly urgent issues. The detection of allergen-specific antibodies, in particular, sIgE and sIgG, is a modern alternative to skin tests due to the safety and efficiency of this method. The use of allergen microarrays to detect tens to hundreds of allergen-specific antibodies in less than 0.1 mL of blood serum enables the transition to a deeply personalized approach in the diagnosis of these diseases while reducing the invasiveness and increasing the informativeness of analysis. This review discusses the technological approaches underlying the development of allergen microarrays and other protein microarrays, including the methods of selection of the microarray substrates and matrices for protein molecule immobilization, the obtainment of allergens, and the use of different types of optical labels for increasing the sensitivity and specificity of the detection of allergen-specific antibodies.
近几十年来,过敏性疾病的发病率急剧上升,其原因包括接触环境诱因的机会越来越多、饮食习惯的改变、合并症以及药物使用的增加。在这种情况下,对各种过敏原的过敏性进行多重诊断以及监测过敏性疾病的治疗效果就成了尤为紧迫的问题。检测过敏原特异性抗体,特别是 sIgE 和 sIgG,因其安全高效而成为皮试的现代替代方法。使用过敏原微阵列可在不到 0.1 毫升的血清中检测数十种至数百种过敏原特异性抗体,从而在诊断这些疾病时过渡到深度个性化的方法,同时降低侵入性并增加分析的信息量。本综述讨论了开发过敏原芯片和其他蛋白质芯片的基本技术方法,包括芯片基底和蛋白质分子固定基质的选择方法、过敏原的获取以及使用不同类型的光学标签来提高过敏原特异性抗体检测的灵敏度和特异性。
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引用次数: 0
Capacitive Neuromodulation via Material-Based Passive Interaction: Efficacy in Motor Function Improvement in Parkinson Disease 通过基于材料的被动交互进行电容性神经调制:改善帕金森病患者运动功能的功效
Pub Date : 2024-07-20 DOI: 10.3390/bios14070354
Fabrizio D’Errico, Francesco Serio, Gianluigi Carioni
A non-invasive and non-pharmacological approach is evaluated for the proprioceptive and postural improvement of PD subjects. The authors evaluated the effectiveness of a class I medical device according to EU regulation 745/2017 designed to develop the mechanism of action based on the modulation of action potentials, which occurs in prevalent pathways of the afferent peripheral nervous system efferent in subjects with spasticity. The present observational study, structured in a double-blind randomized manner, therefore, had the main aim of evaluating the ability of the device to improve on the motor and proprioceptive function of PD patients. This study was based on the instrumented gait analysis performed according to the Timed Up and Go (TUG) test procedure, as well as using a fall risk assessment in accordance with the Berg Balance Scale (BBS) procedures. This study involved 25 participants in the active group (no placebo) and 25 in the non-active group (placebo), the latter to whom non-functional devices were applied, but in every respect identical to the functional devices applied to the 25 patients in the no placebo group. Data analysis was conducted using statistical methodologies for statistics, the statistical significance of the results for the observed samples and the interdependence between the measured variables. The study of the mechanism of action based on the remodulation of action potentials was preliminary conducted through numerical modeling of the Hodgkin–Huxley axon, modified by introducing the influence of the capacitive device applied in clinical tests into the validated model to target the dielectric properties of materials constituting the passive sensor. The use of the neuromodulation device promises observable improvements in motor function among PD patients, including increased limb mobility and greater postural stability.
该研究评估了一种非侵入性和非药物疗法,用于改善帕金森病患者的本体感觉和姿势。作者根据欧盟第 745/2017 号法规评估了一种 I 类医疗设备的有效性,该设备旨在开发基于动作电位调节的作用机制,这种作用机制发生在痉挛受试者外周神经系统传入传出的流行通路中。因此,本观察性研究以双盲随机方式进行,主要目的是评估该设备改善脊髓灰质炎患者运动和本体感觉功能的能力。这项研究基于仪器步态分析,按照定时起立和前进(TUG)测试程序进行,并使用伯格平衡量表(BBS)程序进行跌倒风险评估。这项研究涉及积极组(无安慰剂)和非积极组(安慰剂)各 25 名参与者,后者使用的是非功能性装置,但在各方面都与无安慰剂组 25 名患者使用的功能性装置相同。数据分析采用了统计方法,用于统计观察样本结果的统计意义以及测量变量之间的相互依存关系。通过对霍奇金-赫胥黎轴突进行数值建模,对基于动作电位重塑的作用机制进行了初步研究,并将临床试验中应用的电容式装置的影响引入到验证模型中,针对构成被动传感器的材料的介电特性进行了修改。神经调控装置的使用有望明显改善帕金森病患者的运动功能,包括增强肢体活动能力和姿势稳定性。
{"title":"Capacitive Neuromodulation via Material-Based Passive Interaction: Efficacy in Motor Function Improvement in Parkinson Disease","authors":"Fabrizio D’Errico, Francesco Serio, Gianluigi Carioni","doi":"10.3390/bios14070354","DOIUrl":"https://doi.org/10.3390/bios14070354","url":null,"abstract":"A non-invasive and non-pharmacological approach is evaluated for the proprioceptive and postural improvement of PD subjects. The authors evaluated the effectiveness of a class I medical device according to EU regulation 745/2017 designed to develop the mechanism of action based on the modulation of action potentials, which occurs in prevalent pathways of the afferent peripheral nervous system efferent in subjects with spasticity. The present observational study, structured in a double-blind randomized manner, therefore, had the main aim of evaluating the ability of the device to improve on the motor and proprioceptive function of PD patients. This study was based on the instrumented gait analysis performed according to the Timed Up and Go (TUG) test procedure, as well as using a fall risk assessment in accordance with the Berg Balance Scale (BBS) procedures. This study involved 25 participants in the active group (no placebo) and 25 in the non-active group (placebo), the latter to whom non-functional devices were applied, but in every respect identical to the functional devices applied to the 25 patients in the no placebo group. Data analysis was conducted using statistical methodologies for statistics, the statistical significance of the results for the observed samples and the interdependence between the measured variables. The study of the mechanism of action based on the remodulation of action potentials was preliminary conducted through numerical modeling of the Hodgkin–Huxley axon, modified by introducing the influence of the capacitive device applied in clinical tests into the validated model to target the dielectric properties of materials constituting the passive sensor. The use of the neuromodulation device promises observable improvements in motor function among PD patients, including increased limb mobility and greater postural stability.","PeriodicalId":100185,"journal":{"name":"Biosensors","volume":"84 1","pages":""},"PeriodicalIF":0.0,"publicationDate":"2024-07-20","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141742168","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
引用次数: 0
Development of a “Signal-On” Fluorescent Aptasensor for Highly Selective and Sensitive Detection of ZEN in Cereal Products Using Nitrogen-Doped Carbon Dots Based on the Inner Filter Effect 利用基于内滤光器效应的掺氮碳点开发用于高选择性和高灵敏度检测谷物产品中 ZEN 的 "信号灯 "荧光光度传感器
Pub Date : 2024-07-17 DOI: 10.3390/bios14070347
Qi Sun, Yuting Zhou, Miaomiao Ma, Fuyan Zhang, Shuang Li, Zhuoer Chen, Yu Fang, Tao Le, Fuguo Xing
This study aimed to develop a novel fluorescent aptasensor for the quantitative detection of zearalenone (ZEN), addressing the limitations of conventional detection techniques in terms of speed, sensitivity, and ease of use. Nitrogen-doped carbon dots (N-CDs) were synthesized via the hydrothermal method, resulting in spherical particles with a diameter of 3.25 nm. These N-CDs demonstrated high water solubility and emitted a bright blue light at 440 nm when excited at 355 nm. The fluorescence of N-CDs was quenched by dispersed gold nanoparticles (AuNPs) through the inner filter effect, while aggregated AuNPs induced by NaCl did not affect the fluorescence of N-CDs. The aptamer could protect AuNPs from NaCl-induced aggregation, but the presence of ZEN weakened this protective effect. Based on this principle, optimal conditions for ZEN detection included 57 mM NaCl, 12.5 nM aptamer concentration, incubation of AuNPs with NaCl for 15 min in Tris-EDTA(TE) buffer, and incubation of aptamer with ZEN and NaCl for 30 min. Under these optimized conditions, the “signal-on” fluorescent aptasensor for ZEN detection showed a linear range of 0.25 to 200 ng/mL with a low detection limit of 0.0875 ng/mL. Furthermore, the developed aptasensor exhibited excellent specificity and could rapidly detect ZEN in corn flour samples or corn oil, achieving satisfactory recovery rates ranging from 84.7% to 108.6%. Therefore, this study presents an economical, convenient, sensitive, and rapid method for accurately quantifying ZEN in cereal products.
本研究旨在开发一种用于定量检测玉米赤霉烯酮(ZEN)的新型荧光传感器,以解决传统检测技术在速度、灵敏度和易用性方面的局限性。通过水热法合成了掺氮碳点(N-CDs),形成了直径为 3.25 纳米的球形颗粒。这些 N-CDs 具有很高的水溶性,在 355 纳米波长下激发时会在 440 纳米波长处发出明亮的蓝光。分散的金纳米粒子(AuNPs)通过内滤效应淬灭了 N-CDs 的荧光,而 NaCl 诱导的 AuNPs 聚合不会影响 N-CDs 的荧光。适配体可以保护 AuNPs 免受 NaCl 诱导的聚集,但 ZEN 的存在削弱了这种保护作用。根据这一原理,ZEN 检测的最佳条件包括:57 mM NaCl、12.5 nM 的适配体浓度、AuNPs 在 Tris-EDTA(TE)缓冲液中与 NaCl 一起孵育 15 分钟,以及适配体与 ZEN 和 NaCl 一起孵育 30 分钟。在这些优化条件下,用于检测 ZEN 的 "signal-on "荧光灵敏传感器的线性范围为 0.25 至 200 ng/mL,检测限低至 0.0875 ng/mL。此外,所开发的适配传感器具有极佳的特异性,可快速检测玉米粉或玉米油中的 ZEN,回收率在 84.7% 至 108.6% 之间,令人满意。因此,本研究提出了一种经济、方便、灵敏、快速的方法来准确定量谷物产品中的 ZEN。
{"title":"Development of a “Signal-On” Fluorescent Aptasensor for Highly Selective and Sensitive Detection of ZEN in Cereal Products Using Nitrogen-Doped Carbon Dots Based on the Inner Filter Effect","authors":"Qi Sun, Yuting Zhou, Miaomiao Ma, Fuyan Zhang, Shuang Li, Zhuoer Chen, Yu Fang, Tao Le, Fuguo Xing","doi":"10.3390/bios14070347","DOIUrl":"https://doi.org/10.3390/bios14070347","url":null,"abstract":"This study aimed to develop a novel fluorescent aptasensor for the quantitative detection of zearalenone (ZEN), addressing the limitations of conventional detection techniques in terms of speed, sensitivity, and ease of use. Nitrogen-doped carbon dots (N-CDs) were synthesized via the hydrothermal method, resulting in spherical particles with a diameter of 3.25 nm. These N-CDs demonstrated high water solubility and emitted a bright blue light at 440 nm when excited at 355 nm. The fluorescence of N-CDs was quenched by dispersed gold nanoparticles (AuNPs) through the inner filter effect, while aggregated AuNPs induced by NaCl did not affect the fluorescence of N-CDs. The aptamer could protect AuNPs from NaCl-induced aggregation, but the presence of ZEN weakened this protective effect. Based on this principle, optimal conditions for ZEN detection included 57 mM NaCl, 12.5 nM aptamer concentration, incubation of AuNPs with NaCl for 15 min in Tris-EDTA(TE) buffer, and incubation of aptamer with ZEN and NaCl for 30 min. Under these optimized conditions, the “signal-on” fluorescent aptasensor for ZEN detection showed a linear range of 0.25 to 200 ng/mL with a low detection limit of 0.0875 ng/mL. Furthermore, the developed aptasensor exhibited excellent specificity and could rapidly detect ZEN in corn flour samples or corn oil, achieving satisfactory recovery rates ranging from 84.7% to 108.6%. Therefore, this study presents an economical, convenient, sensitive, and rapid method for accurately quantifying ZEN in cereal products.","PeriodicalId":100185,"journal":{"name":"Biosensors","volume":"151 1","pages":""},"PeriodicalIF":0.0,"publicationDate":"2024-07-17","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141719114","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
引用次数: 0
Enhanced Point-of-Care SARS-CoV-2 Detection: Integrating RT-LAMP with Microscanning 加强护理点 SARS-CoV-2 检测:RT-LAMP 与显微扫描相结合
Pub Date : 2024-07-17 DOI: 10.3390/bios14070348
Minkyeong Choi, Eunji Lee, Seoyeon Park, Chae-Seung Lim, Woong-Sik Jang
The COVID-19 pandemic has highlighted the urgent need for rapid and accurate diagnostic methods for various infectious diseases, including SARS-CoV-2. Traditional RT-PCR methods, while highly sensitive and specific, require complex equipment and skilled personnel. In response, we developed an integrated RT-LAMP-MS assay, which combines rapid reverse transcription loop-mediated isothermal amplification (RT-LAMP) with microscanning (MS) technology for detecting SARS-CoV-2. The assay uses magnesium pyrophosphate formed during LAMP amplification as a visual marker, allowing direct observation via microscopy without the need for additional chemical indicators or probes. For the SARS-CoV-2/IC RT-LAMP-MS assay, the sample-LAMP reagent mixture was added to a microchip with SARS-CoV-2 primers and internal controls, then incubated at 62 °C for 30 min in a heat block, followed by amplification analysis using a microscanner. In clinical tests, the RT-LAMP-MS assay showed 99% sensitivity and 100% specificity, which is identical to the RT-LAMP results and comparable to the commercial AllplexTM SARS-CoV-2 assay results. Additionally, the limit of detection (LOD) was determined to be 10-¹ PFU mL⁻¹ (dynamic range: 103~10−¹ PFU mL⁻¹). The assay delivers results in 30 min, uses low-cost equipment, and demonstrates 100% reproducibility in repeated tests, making it suitable for point-of-care use in resource-limited settings.
COVID-19 大流行凸显了对包括 SARS-CoV-2 在内的各种传染病的快速准确诊断方法的迫切需求。传统的 RT-PCR 方法虽然灵敏度高、特异性强,但需要复杂的设备和熟练的人员。为此,我们开发了一种 RT-LAMP-MS 集成测定法,它将快速反转录环介导等温扩增(RT-LAMP)与显微扫描(MS)技术相结合,用于检测 SARS-CoV-2。该检测方法使用 LAMP 扩增过程中形成的焦磷酸镁作为视觉标记,可通过显微镜直接观察,而无需额外的化学指示剂或探针。在 SARS-CoV-2/IC RT-LAMP-MS 检测中,将样品-LAMP 试剂混合物与 SARS-CoV-2 引物和内部对照一起添加到微芯片中,然后在加热块中于 62 °C 温育 30 分钟,接着使用显微镜进行扩增分析。在临床试验中,RT-LAMP-MS 检测法的灵敏度为 99%,特异性为 100%,与 RT-LAMP 检测法的结果相同,与商用 AllplexTM SARS-CoV-2 检测法的结果相当。此外,检测限(LOD)被确定为 10-¹ PFU mL-¹(动态范围:103~10-¹ PFU mL-¹)。该检测方法可在 30 分钟内得出结果,使用的设备成本低,重复检测的重现性达到 100%,因此适合在资源有限的环境中进行床旁检测。
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引用次数: 0
Beta-Barrel Nanopores as Diagnostic Sensors: An Engineering Perspective 作为诊断传感器的贝塔管纳米孔:工程学视角
Pub Date : 2024-07-16 DOI: 10.3390/bios14070345
Rani Wiswedel, Anh Thi Ngoc Bui, Jinhyung Kim, Mi-Kyung Lee
Biological nanopores are ultrasensitive and highly attractive platforms for disease diagnostics, including the sequencing of viral and microbial genes and the detection of biomarkers and pathogens. To utilize biological nanopores as diagnostic sensors, they have been engineered through various methods resulting in the accurate and highly sensitive detection of biomarkers and disease-related biomolecules. Among diverse biological nanopores, the β-barrel-containing nanopores have advantages in nanopore engineering because of their robust structure, making them well-suited for modifications. In this review, we highlight the engineering approaches for β-barrel-containing nanopores used in single-molecule sensing for applications in early diagnosis and prognosis. In the highlighted studies, β-barrel nanopores can be modified by genetic mutation to change the structure; alter charge distributions; or add enzymes, aptamers, and protein probes to enhance sensitivity and accuracy. Furthermore, this review discusses challenges and future perspectives for advancing nanopore-based diagnostic sensors.
生物纳米孔是超灵敏和极具吸引力的疾病诊断平台,包括病毒和微生物基因测序以及生物标记物和病原体检测。为了将生物纳米孔用作诊断传感器,人们通过各种方法对其进行了改造,从而实现了对生物标记物和疾病相关生物分子的精确、高灵敏度检测。在各种生物纳米孔中,含β管的纳米孔在纳米孔工程中具有优势,因为其结构坚固,非常适合改性。在这篇综述中,我们重点介绍了用于单分子传感的含β管纳米孔的工程方法,这些方法可应用于早期诊断和预后。在重点研究中,β 管纳米孔可以通过基因突变来改变结构;改变电荷分布;或添加酶、适配体和蛋白质探针,以提高灵敏度和准确性。此外,本综述还讨论了推进基于纳米孔的诊断传感器所面临的挑战和未来展望。
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引用次数: 0
Aptamer Based on Silver Nanoparticle-Modified Flexible Carbon Ink Printed Electrode for the Electrochemical Detection of Chikungunya Virus 用于电化学检测基孔肯雅病毒的基于银纳米粒子修饰的柔性碳墨印刷电极的适配体
Pub Date : 2024-07-16 DOI: 10.3390/bios14070344
Pradakshina Sharma, Mohd. Rahil Hasan, Ubaid Mushtaq Naikoo, Shaheen Khatoon, R. Pilloton, Jagriti Narang
Medical devices have progressed from their initial bulky forms to smart devices. However, their rigidity hampers their seamless integration into everyday life. The fields of stretchable, textile, and flexible electronics are emerging research areas with the potential to drive significant technological progress. This research presents a laboratory-based technique to produce highly sensitive and flexible biosensors for detecting the chikungunya virus. These biosensors are based on 0D nanomaterials and demonstrate significant advancements in voltammetry. The electrochemical platform was created utilizing the stencil printing (StPE) technique. Adapting the biosensor setup involved the selection of aptamer as the biorecognition element bound with silver nanoparticles (AgNPs). This biosensor was employed in the voltammetric identification of the Chikungunya virus antigen (CHIKV-Ag) within a solution containing 0.5 mM potassium ferro/ferri cyanide, a redox pair. The biosensor was employed to evaluate CHIKV-Ag within a human serum sample. It demonstrated a linear detection span ranging from 0.1 ng/mL to 1 μg/mL, with a detection limit of 0.1 ng/mL for CHIKV-Ag. The proposed approach, due to its flexibility in production and the electrocatalytic attributes displayed by the zero-dimensional nanostructure, presents innovative opportunities for cost-effective and tailored aptamer-based bioelectronics, thereby broadening the scope of this domain.
医疗设备已从最初的笨重形态发展为智能设备。然而,它们的刚性阻碍了它们与日常生活的无缝融合。可拉伸、纺织品和柔性电子领域是新兴的研究领域,有可能推动重大技术进步。本研究介绍了一种基于实验室的技术,用于生产检测基孔肯雅病毒的高灵敏度柔性生物传感器。这些生物传感器基于 0D 纳米材料,展示了伏安法的重大进展。电化学平台是利用模板印刷(StPE)技术制作的。调整生物传感器的设置涉及选择与银纳米粒子(AgNPs)结合的适配体作为生物识别元素。该生物传感器被用于在含有 0.5 mM 铁氰化钾(氧化还原对)的溶液中对基孔肯雅病毒抗原(CHIKV-Ag)进行伏安识别。该生物传感器用于评估人体血清样本中的 CHIKV-Ag。它的线性检测范围为 0.1 ng/mL 至 1 μg/mL,CHIKV-Ag 的检测限为 0.1 ng/mL。由于其生产的灵活性和零维纳米结构所显示的电催化特性,所提出的方法为基于贴体的生物电子学提供了具有成本效益和量身定制的创新机会,从而拓宽了这一领域的范围。
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引用次数: 0
Ultrasonic Sensor: A Fast and Non-Destructive System to Measure the Viscosity and Density of Molecular Fluids 超声波传感器:测量分子流体粘度和密度的快速无损系统
Pub Date : 2024-07-16 DOI: 10.3390/bios14070346
Romina Muñoz, Juan-Francisco Fuentealba, Sebastián Michea, Paula A. Santana, Juan Ignacio Martinez, Nathalie Casanova-Morales, Vicente Salinas-Barrera
This study presents the design and development of an ultrasonic sensor as a fundamental tool for characterizing the properties of fluids and biofluids. The analysis primarily focuses on measuring the electrical parameters of the system, which correlate with the density and viscosity of the solutions, in sample volumes of microliters and with high temporal resolution (up to 1 data point per second). The use of this sensor allows the fast and non-destructive evaluation of the viscosity and density of fluids deposited on its free surface. The measurements are based on obtaining the impedance versus frequency curve and the phase difference curve (between current and voltage) versus frequency. In this way, characteristic parameters of the transducer, such as the resonance frequency, phase, minimum impedance, and the quality factor of the resonant system, can characterize variations in density and viscosity in the fluid under study. The results obtained revealed the sensor’s ability to identify two parameters sensitive to viscosity and two parameters sensitive to density. As a proof of concept, the unfolding of the bovine albumin protein was studied, resulting in a curve that reflects its unfolding kinetics in the presence of urea.
本研究介绍了超声波传感器的设计和开发,它是表征流体和生物流体特性的基本工具。分析的主要重点是测量系统的电参数,这些参数与溶液的密度和粘度相关,样品量为微升,时间分辨率高(每秒最多 1 个数据点)。使用这种传感器可以快速、无损地评估沉积在其自由表面上的液体的粘度和密度。测量的基础是获得阻抗随频率变化的曲线以及相位差(电流和电压之间)随频率变化的曲线。这样,传感器的特征参数,如共振频率、相位、最小阻抗和共振系统的品质因数,就可以描述所研究流体的密度和粘度变化。研究结果表明,传感器能够识别对粘度敏感的两个参数和对密度敏感的两个参数。作为概念验证,对牛白蛋白的展开进行了研究,得出了反映其在尿素存在下展开动力学的曲线。
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引用次数: 0
期刊
Biosensors
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