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Paralytic Shellfish Toxin Extraction from Bivalve Meat for Analysis Using Potentiometric Chemical Sensors. 从双壳贝肉中提取麻痹性贝类毒素,利用电位化学传感器进行分析。
IF 4.9 3区 工程技术 Q1 CHEMISTRY, ANALYTICAL Pub Date : 2024-10-08 DOI: 10.3390/bios14100487
Ana Filipa R Cerqueira, Catarina Moreirinha, Mariana Raposo, Maria Teresa S R Gomes, Sara T Costa, Maria João Botelho, Alisa Rudnitskaya

A simple and reliable methodology for the detection of paralytic shellfish toxins (PSTs) in bivalve tissues using potentiometric chemical sensors was developed. Five methods of PST extraction from mussel and oyster tissues were evaluated, including the AOAC-recommended method, which served as the reference. The main objective was to minimize the matrix effect of the extracts on the sensors' responses and ensure efficient toxin recovery. Extraction procedures using acetic acid with heating and water yielded the highest responses from the potentiometric chemical sensors to PSTs. The highest recovery of PSTs from bivalve tissues was achieved with extraction using acetic acid and heating. Further extract purification, which is indispensable for liquid chromatography with fluorometric detection (LC-FLD) analysis, was found to be unnecessary for analysis with chemical sensors. While water extraction can also be used as a rapid and simple PST extraction method, the lower recoveries should be considered when interpreting the results. Further research is needed to identify the compounds remaining in the extracts that cause a decrease in sensor responses and to develop procedures for their elimination.

利用电位化学传感器开发了一种简单可靠的方法来检测双壳贝类组织中的麻痹性贝类毒素(PST)。评估了从贻贝和牡蛎组织中提取 PST 的五种方法,包括 AOAC 推荐的参考方法。主要目的是尽量减少提取物对传感器响应的基质效应,并确保有效的毒素回收。使用醋酸加热和水进行提取的程序使电位化学传感器对 PST 的反应最高。使用醋酸和加热萃取法从双壳类组织中回收的 PST 最高。进一步的提取纯化对于液相色谱-荧光检测(LC-FLD)分析是必不可少的,但对于使用化学传感器进行分析则没有必要。虽然水提取也可作为一种快速、简单的 PST 提取方法,但在解释结果时应考虑到较低的回收率。需要进一步研究,以确定萃取物中残留的导致传感器响应降低的化合物,并制定消除这些化合物的程序。
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引用次数: 0
A Wireless and Wearable Multimodal Sensor to Non-Invasively Monitor Transabdominal Placental Oxygen Saturation and Maternal Physiological Signals. 用于无创监测经腹胎盘血氧饱和度和产妇生理信号的无线可穿戴多模态传感器。
IF 4.9 3区 工程技术 Q1 CHEMISTRY, ANALYTICAL Pub Date : 2024-10-07 DOI: 10.3390/bios14100481
Thien Nguyen, Soongho Park, Asma Sodager, Jinho Park, Dahiana M Gallo, Guoyang Luo, Roberto Romero, Amir Gandjbakhche

Poor placental development and placental defects can lead to adverse pregnancy outcomes such as pre-eclampsia, fetal growth restriction, and stillbirth. This study introduces two sensors, which use a near-infrared spectroscopy (NIRS) technique to measure placental oxygen saturation transabdominally. The first one, an NIRS sensor, is a wearable device consisting of multiple NIRS channels. The second one, a Multimodal sensor, which is an upgraded version of the NIRS sensor, is a wireless and wearable device, integrating a motion sensor and multiple NIRS channels. A pilot clinical study was conducted to assess the feasibility of the two sensors in measuring transabdominal placental oxygenation in 36 pregnant women (n = 12 for the NIRS sensor and n = 24 for the Multimodal sensor). Among these subjects, 4 participants had an uncomplicated pregnancy, and 32 patients had either maternal pre-existing conditions/complications, neonatal complications, and/or placental pathologic abnormalities. The study results indicate that the patients with maternal complicated conditions (69.5 ± 5.4%), placental pathologic abnormalities (69.4 ± 4.9%), and neonatal complications (68.0 ± 5.1%) had statistically significantly lower transabdominal placental oxygenation levels than those with an uncomplicated pregnancy (76.0 ± 4.4%) (F (3,104) = 6.6, p = 0.0004). Additionally, this study shows the capability of the Multimodal sensor in detecting the maternal heart rate and respiratory rate, fetal movements, and uterine contractions. These findings demonstrate the feasibility of the two sensors in the real-time continuous monitoring of transabdominal placental oxygenation to detect at-risk pregnancies and guide timely clinical interventions, thereby improving pregnancy outcomes.

胎盘发育不良和胎盘缺陷可导致不良妊娠结局,如先兆子痫、胎儿生长受限和死胎。本研究介绍了两种利用近红外光谱(NIRS)技术经腹测量胎盘血氧饱和度的传感器。第一种是近红外传感器,是一种由多个近红外通道组成的可穿戴设备。第二种是多模态传感器,它是近红外传感器的升级版,是一种无线可穿戴设备,集成了运动传感器和多个近红外光谱通道。为了评估这两种传感器测量经腹胎盘氧合的可行性,我们对 36 名孕妇(近红外传感器为 12 人,多模态传感器为 24 人)进行了试点临床研究。在这些受试者中,4 名受试者为无并发症妊娠,32 名患者患有母体原有疾病/并发症、新生儿并发症和/或胎盘病理异常。研究结果表明,与无并发症妊娠的患者(76.0 ± 4.4%)相比,患有产妇并发症(69.5 ± 5.4%)、胎盘病理异常(69.4 ± 4.9%)和新生儿并发症(68.0 ± 5.1%)的患者经腹胎盘氧合水平明显较低(F (3,104) = 6.6,P = 0.0004)。此外,该研究还显示了多模态传感器检测母体心率和呼吸频率、胎动和子宫收缩的能力。这些研究结果证明了这两种传感器在实时连续监测经腹胎盘氧合方面的可行性,可用于检测高危妊娠并指导及时的临床干预,从而改善妊娠结局。
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引用次数: 0
A Low-Cost, Portable, Multi-Cancer Screening Device Based on a Ratio Fluorometry and Signal Correlation Technique. 基于比率荧光测定法和信号相关技术的低成本、便携式、多种癌症筛查设备。
IF 4.9 3区 工程技术 Q1 CHEMISTRY, ANALYTICAL Pub Date : 2024-10-07 DOI: 10.3390/bios14100482
Abdulaziz S Alghamdi, Rabah W Aldhaheri

The autofluorescence of erythrocyte porphyrins has emerged as a potential method for multi-cancer early detection (MCED). With this method's dependence on research-grade spectrofluorometers, significant improvements in instrumentation are necessary to translate its potential into clinical practice, as with any promising medical technology. To fill this gap, in this paper, we present an automated ratio porphyrin analyzer for cancer screening (ARPA-CS), a low-cost, portable, and automated instrument for MCED via the ratio fluorometry of porphyrins. The ARPA-CS aims to facilitate cancer screening in an inexpensive, rapid, non-invasive, and reasonably accurate manner for use in primary clinics or at point of care. To accomplish this, the ARPA-CS uses an ultraviolet-excited optical apparatus for ratio fluorometry that features two photodetectors for detection at 590 and 630 nm. Additionally, it incorporates a synchronous detector for the precision measurement of signals based on the Walsh-ordered Walsh-Hadamard transform (WHT)w and circular shift. To estimate its single-photodetector capability, we established a linear calibration curve for the ARBA-CS exceeding four orders of magnitude with a linearity of up to 0.992 and a low detection limit of 0.296 µg/mL for riboflavin. The ARPA-CS also exhibited excellent repeatability (0.21%) and stability (0.60%). Moreover, the ratio fluorometry of three serially diluted standard solutions of riboflavin yielded a ratio of 0.4, which agrees with that expected based on the known emission spectra of riboflavin. Additionally, the ratio fluorometry of the porphyrin solution yielded a ratio of 49.82, which was ascribed to the predominant concentration of protoporphyrin IX in the brown eggshells, as confirmed in several studies. This study validates this instrument for the ratio fluorometry of porphyrins as a biomarker for MCED. Nevertheless, large and well-designed clinical trials are necessary to further elaborate more on this matter.

红细胞卟啉的自发荧光已成为一种潜在的多癌症早期检测(MCED)方法。由于这种方法依赖于研究级的分光荧光仪,因此要将其潜力转化为临床实践,就必须对仪器进行重大改进,就像任何有前途的医疗技术一样。为了填补这一空白,我们在本文中介绍了用于癌症筛查的自动卟啉比值分析仪(ARPA-CS),这是一种通过卟啉比值荧光测定法进行 MCED 的低成本、便携式和自动化仪器。ARPA-CS 旨在以廉价、快速、无创和合理准确的方式促进癌症筛查,供基层诊所或医疗点使用。为实现这一目标,ARPA-CS 使用紫外线激发光学仪器进行比率荧光测定,该仪器有两个光电探测器,分别在 590 和 630 纳米波长进行检测。此外,它还集成了一个同步探测器,用于根据沃尔什有序沃尔什-哈达玛德变换 (WHT)w 和圆周偏移精确测量信号。为了评估其单光电探测器的能力,我们为 ARBA-CS 建立了一条线性校准曲线,其线性度超过四个数量级,线性度高达 0.992,核黄素的检测限低至 0.296 µg/mL。ARPA-CS 还具有出色的重复性(0.21%)和稳定性(0.60%)。此外,对核黄素的三种连续稀释的标准溶液进行比率荧光测定,得出的比率为 0.4,这与根据已知的核黄素发射光谱预期的比率一致。此外,对卟啉溶液进行比率荧光测定得出的比率为 49.82,这是因为棕色蛋壳中主要含有原卟啉 IX,这一点已在多项研究中得到证实。这项研究验证了该仪器可将卟啉比率荧光测定法作为 MCED 的生物标志物。不过,还需要进行大规模和设计良好的临床试验,以进一步阐明这一问题。
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引用次数: 0
Principles and Applications of ZnO Nanomaterials in Optical Biosensors and ZnO Nanomaterial-Enhanced Biodetection. 氧化锌纳米材料在光学生物传感器和氧化锌纳米材料增强生物检测中的原理和应用。
IF 4.9 3区 工程技术 Q1 CHEMISTRY, ANALYTICAL Pub Date : 2024-10-06 DOI: 10.3390/bios14100480
Marion Ryan C Sytu, Jong-In Hahm

Significant research accomplishments have been made so far for the development and application of ZnO nanomaterials in enhanced optical biodetection. The unparalleled optical properties of ZnO nanomaterials and their reduced dimensionality have been successfully exploited to push the limits of conventional optical biosensors and optical biodetection platforms for a wide range of bioanalytes. ZnO nanomaterial-enabled advancements in optical biosensors have been demonstrated to improve key sensor performance characteristics such as the limit of detection and dynamic range. In addition, all nanomaterial forms of ZnO, ranging from 0-dimensional (0D) and 1D to 2D nanostructures, have been proven to be useful, ensuring their versatile fabrication into functional biosensors. The employment of ZnO as an essential biosensing element has been assessed not only for ensembles but also for individual nanomaterials, which is advantageous for the realization of high miniaturization and minimal invasiveness in biosensors and biodevices. Moreover, the nanomaterials' incorporations into biosensors have been shown to be useful and functional for a variety of optical detection modes, such as absorption, colorimetry, fluorescence, near-band-edge emission, deep-level emission, chemiluminescence, surface evanescent wave, whispering gallery mode, lossy-mode resonance, surface plasmon resonance, and surface-enhanced Raman scattering. The detection capabilities of these ZnO nanomaterial-based optical biosensors demonstrated so far are highly encouraging and, in some cases, permit quantitative analyses of ultra-trace level bioanalytes that cannot be measured by other means. Hence, steady research endeavors are expected in this burgeoning field, whose scientific and technological impacts will grow immensely in the future. This review provides a timely and much needed review of the research efforts made in the field of ZnO nanomaterial-based optical biosensors in a comprehensive and systematic manner. The topical discussions in this review are organized by the different modes of optical detection listed above and further grouped by the dimensionality of the ZnO nanostructures used in biosensors. Following an overview of a given optical detection mode, the unique properties of ZnO nanomaterials critical to enhanced biodetection are presented in detail. Subsequently, specific biosensing applications of ZnO nanomaterials are discussed for ~40 different bioanalytes, and the important roles that the ZnO nanomaterials play in bioanalyte detection are also identified.

迄今为止,在增强型光学生物检测中开发和应用氧化锌纳米材料的研究取得了重大成就。人们成功地利用氧化锌纳米材料无与伦比的光学特性及其减小的尺寸,突破了传统光学生物传感器和光学生物检测平台对多种生物分析物的检测极限。氧化锌纳米材料在光学生物传感器中的应用已证明可改善传感器的主要性能特征,如检测极限和动态范围。此外,氧化锌的所有纳米材料形式,从 0 维(0D)、1D 到 2D 纳米结构,都已被证明是有用的,从而确保了其在功能性生物传感器中的多功能性。氧化锌作为一种重要的生物传感元素,不仅在组合方面,而且在单个纳米材料方面都得到了评估,这有利于实现生物传感器和生物设备的高度微型化和微创化。此外,将纳米材料融入生物传感器已被证明可用于多种光学检测模式,如吸收、比色、荧光、近带边发射、深层发射、化学发光、表面蒸发波、耳语画廊模式、有损模式共振、表面等离子体共振和表面增强拉曼散射。迄今为止,这些基于氧化锌纳米材料的光学生物传感器的检测能力非常令人鼓舞,在某些情况下,可以对其他方法无法测量的超痕量生物分析物进行定量分析。因此,预计这一新兴领域的研究工作将持续进行,其科学和技术影响将在未来大幅增长。本综述以全面、系统的方式对基于氧化锌纳米材料的光学生物传感器领域的研究工作进行了及时和急需的回顾。本综述中的专题讨论按照上述不同的光学检测模式组织,并根据生物传感器中使用的氧化锌纳米结构的尺寸进一步分组。在概述了特定光学检测模式之后,详细介绍了对增强生物检测至关重要的氧化锌纳米材料的独特性能。随后,针对约 40 种不同的生物分析物讨论了氧化锌纳米材料的具体生物传感应用,并确定了氧化锌纳米材料在生物分析物检测中的重要作用。
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引用次数: 0
Comparison of Survivin Determination by Surface-Enhanced Fluorescence and Raman Spectroscopy on Nanostructured Silver Substrates. 通过表面增强荧光和拉曼光谱在纳米结构银基底上测定 Survivin 的比较。
IF 4.9 3区 工程技术 Q1 CHEMISTRY, ANALYTICAL Pub Date : 2024-10-06 DOI: 10.3390/bios14100479
Georgia Geka, Anastasia Kanioura, Ioannis Kochylas, Vlassis Likodimos, Spiros Gardelis, Anastasios Dimitriou, Nikolaos Papanikolaou, Anastasios Economou, Sotirios Kakabakos, Panagiota Petrou

Survivin belongs to a family of proteins that promote cellular proliferation and inhibit cellular apoptosis. Its overexpression in various cancer types has led to its recognition as an important marker for cancer diagnosis and treatment. In this work, we compare two approaches for the immunochemical detection of survivin through surface-enhanced fluorescence or Raman spectroscopy using surfaces with nanowires decorated with silver nanoparticles in the form of dendrites or aggregates as immunoassays substrates. In both substrates, a two-step non-competitive immunoassay was developed using a pair of specific monoclonal antibodies, one for detection and the other for capture. The detection antibody was biotinylated and combined with streptavidin labeled with rhodamine for the detection of surface-enhanced fluorescence, while, for the detection via Raman spectroscopy, streptavidin labeled with peroxidase was used and the signal was obtained after the application of 3,3',5,5'-tetramethylbenzidine (TMB) precipitating substrate. It was found that the substrate with the silver dendrites provided higher fluorescence signal intensity compared to the substrate with the silver aggregates, while the opposite was observed for the Raman signal. Thus, the best substrate was used for each detection method. A detection limit of 12.5 pg/mL was achieved with both detection approaches along with a linear dynamic range up to 500 pg/mL, enabling survivin determination in human serum samples from both healthy and ovarian cancer patients for cancer diagnosis and monitoring purposes.

Survivin 属于促进细胞增殖和抑制细胞凋亡的蛋白质家族。它在各种癌症类型中的过度表达使其被认为是癌症诊断和治疗的重要标志物。在这项工作中,我们比较了两种通过表面增强荧光或拉曼光谱对存活素进行免疫化学检测的方法,这两种方法都使用了以树枝状或聚集体形式的银纳米粒子装饰的纳米线表面作为免疫测定基底。在这两种基底中,都使用了一对特异性单克隆抗体(一种用于检测,另一种用于捕获),开发了一种两步非竞争性免疫测定。检测抗体经生物素化后与标记有罗丹明的链霉亲和素结合,用于检测表面增强荧光;而通过拉曼光谱进行检测时,则使用标记有过氧化物酶的链霉亲和素,并在使用 3,3',5,5'-四甲基联苯胺(TMB)沉淀底物后获得信号。结果发现,与银聚集体基底相比,银树枝状基底提供的荧光信号强度更高,而拉曼信号则相反。因此,每种检测方法都使用了最佳的基底。两种检测方法的检测限均为 12.5 pg/mL,线性动态范围高达 500 pg/mL,因此可以测定健康人和卵巢癌患者血清样本中的存活素,用于癌症诊断和监测。
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引用次数: 0
Infrared Thermography Sensor in the Analysis of Acute Metabolic Stress Response during Race Walking Competition. 分析竞走比赛中急性代谢应激反应的红外热成像传感器
IF 4.9 3区 工程技术 Q1 CHEMISTRY, ANALYTICAL Pub Date : 2024-10-05 DOI: 10.3390/bios14100478
Alessio Cabizosu, Cristian Marín-Pagan, Pedro E Alcaraz, Francisco Javier Martínez-Noguera

Introduction: Due to the possible impact of the thermoregulatory process on sports performance, it is necessary to explore the existing relationships between kinetic, mechanical, and physiological variables. The objective of this study was to evaluate metabolic stress using thermography in the lower limb after the Spanish Championship 2023 walk. Method: A descriptive study was carried out on national and international race walkers before and after the 2023 Spanish Championships. The participants performed different tests within the same circuit. Five walkers completed the long-distance race of 35 km, four walkers completed the middle-distance race of 20 km and finally, two walkers completed the short-distance race of 10 km. Result: Statistically significant changes were observed in the lower limbs of the walkers after completing the test. We observed a decrease in skin temperature in all the anatomical regions analyzed, except for the back of the leg. More specifically, the decrease was in the hip (-1.92 °C: p = 0.004), quadriceps, hamstrings (-1.23 °C: p = 0.002), and tibia (-1.23 °C: p = 0.030). However, in the posterior region of the leg, a significant increase in temperature was observed (+0.50 °C: p = 0.011) following the competition. Discussion and Conclusions: The findings in this descriptive investigation support the notion that thermography may serve as a useful tool in the acute analysis of muscle functional activation and metabolic response in professional marching athletes. Moreover, the results confirmed that the change in skin temperature is the result of a variation in acute metabolic and functional activation in the lower extremities of race walkers during competition, with infrared thermography representing an instrument capable of detecting such a change in a rapid and non-invasive manner.

导言:由于体温调节过程可能对运动成绩产生影响,因此有必要探索运动、机械和生理变量之间的现有关系。本研究的目的是利用热成像技术评估 2023 年西班牙竞走锦标赛后下肢的代谢压力。研究方法在 2023 年西班牙锦标赛前后,对国内和国际竞走运动员进行了描述性研究。参赛者在同一赛道上进行了不同的测试。五名竞走者完成了 35 公里的长距离比赛,四名竞走者完成了 20 公里的中距离比赛,最后两名竞走者完成了 10 公里的短距离比赛。结果完成测试后,步行者的下肢出现了明显的统计学变化。我们观察到,除腿部后侧外,所有分析区域的皮肤温度都有所下降。更具体地说,下降的部位是臀部(-1.92 °C:p = 0.004)、股四头肌、腘绳肌(-1.23 °C:p = 0.002)和胫骨(-1.23 °C:p = 0.030)。然而,在腿部后侧区域,观察到比赛后温度显著升高(+0.50 °C:p = 0.011)。讨论和结论:这项描述性调查的结果支持这样一种观点,即热成像技术可作为一种有用的工具,用于对专业行进运动员的肌肉功能激活和新陈代谢反应进行急性分析。此外,研究结果证实,皮肤温度的变化是竞走运动员在比赛期间下肢急性代谢和功能激活变化的结果,而红外热成像技术是一种能够以快速和非侵入性方式检测这种变化的仪器。
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引用次数: 0
The Evolution of Illicit-Drug Detection: From Conventional Approaches to Cutting-Edge Immunosensors-A Comprehensive Review. 非法药物检测的演变:从传统方法到尖端免疫传感器--全面回顾。
IF 4.9 3区 工程技术 Q1 CHEMISTRY, ANALYTICAL Pub Date : 2024-10-03 DOI: 10.3390/bios14100477
Nigar Anzar, Shariq Suleman, Yashda Singh, Supriya Kumari, Suhel Parvez, Roberto Pilloton, Jagriti Narang

The increasing use of illicit drugs has become a major global concern. Illicit drugs interact with the brain and the body altering an individual's mood and behavior. As the substance-of-abuse (SOA) crisis continues to spread across the world, in order to reduce trafficking and unlawful activity, it is important to use point-of-care devices like biosensors. Currently, there are certain conventional detection methods, which include gas chromatography (GC), mass spectrometry (MS), surface ionization, surface-enhanced Raman spectroscopy (SERS), surface plasmon resonance (SPR), electrochemiluminescence (ECL), high-performance liquid chromatography (HPLC), etc., for the detection of abused drugs. These methods have the advantage of high accuracy and sensitivity but are generally laborious, expensive, and require trained operators, along with high sample requirements, and they are not suitable for on-site drug detection scenarios. As a result, there is an urgent need for point-of-care technologies for a variety of drugs that can replace conventional techniques, such as a biosensor, specifically an immunosensor. An immunosensor is an analytical device that integrates an antibody-based recognition element with a transducer to detect specific molecules (antigens). In an immunosensor, the highly selective antigen-antibody interaction is used to identify and quantify the target analyte. The binding event between the antibody and antigen is converted by the transducer into a measurable signal, such as electrical, optical, or electrochemical, which corresponds to the presence and concentration of the analyte in the sample. This paper provides a comprehensive overview of various illicit drugs, the conventional methods employed for their detection, and the advantages of immunosensors over conventional techniques. It highlights the critical need for on-site detection and explores emerging point-of-care testing methods. The paper also outlines future research goals in this field, emphasizing the potential of advanced technologies to enhance the accuracy, efficiency, and convenience of drug detection.

非法药物的使用日益增多已成为全球关注的一个主要问题。非法药物会与大脑和身体相互作用,改变人的情绪和行为。随着滥用药物(SOA)危机继续在全球蔓延,为了减少贩运和非法活动,使用生物传感器等护理点设备非常重要。目前,用于检测滥用药物的传统检测方法有气相色谱法(GC)、质谱法(MS)、表面电离法、表面增强拉曼光谱法(SERS)、表面等离子体共振法(SPR)、电化学发光法(ECL)、高效液相色谱法(HPLC)等。这些方法具有准确度和灵敏度高的优点,但一般都比较费力、昂贵,需要经过培训的操作人员,而且对样品的要求较高,不适合现场毒品检测的情况。因此,人们迫切需要能够替代传统技术的各种药物护理点技术,如生物传感器,特别是免疫传感器。免疫传感器是一种分析设备,它将基于抗体的识别元件与传感器集成在一起,用于检测特定分子(抗原)。在免疫传感器中,高度选择性的抗原-抗体相互作用被用来识别和量化目标分析物。抗体和抗原之间的结合事件被传感器转换成可测量的信号,如电信号、光学信号或电化学信号,这些信号与样品中分析物的存在和浓度相对应。本文全面概述了各种非法药物、检测非法药物的传统方法以及免疫传感器相对于传统技术的优势。它强调了现场检测的迫切需要,并探讨了新兴的床旁检测方法。论文还概述了该领域未来的研究目标,强调了先进技术在提高毒品检测的准确性、效率和便利性方面的潜力。
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引用次数: 0
A Double-Stranded Aptamer for Highly Sensitive Fluorescent Detection of Glutathione S-Transferases. 用于高灵敏度荧光检测谷胱甘肽 S 转化酶的双链色聚体。
IF 4.9 3区 工程技术 Q1 CHEMISTRY, ANALYTICAL Pub Date : 2024-10-03 DOI: 10.3390/bios14100476
Wei Cui, Suping Li, Jiahao Zeng, Chen Li, Zhaofeng Li, Xiaohong Wen, Suxia Bao, Yang Mei, Xiangxian Meng, Qiuping Guo

Aptamer-based biosensors have been widely constructed and applied to detect diverse targets. Glutathione S-transferase (GST), a pivotal phase II metabolic enzyme, plays a critical role in biotransformation in vivo, and aberrant GST expression is associated with various health risks. Herein, aptamers targeting GST were systematically selected from a randomized single-stranded DNA (ssDNA) library of 79 nucleotides (nt) using a biotinylated GST-immobilized streptavidin agarose (SA) bead SELEX technology. Following rigorous screening across eight rounds, four aptamers with strikingly similar secondary structures emerged. Among these, Seq3 exhibited the highest affinity towards GST and was selected for further optimization. A semi-rational post-SELEX truncation strategy was then employed based on base composition analysis, secondary structure analysis and affinity assessment. This strategy enabled the systematic removal of redundant nucleotides in Seq3 without compromising its affinity, ultimately yielding a truncated aptamer, Seq3-3, which retains its specificity with a compact 39nt length. Building upon Seq3-3, a double-stranded fluorescent aptamer probe was ingeniously designed for the in vitro detection of GST. The detection mechanism hinges on the competitive displacement of the complementary chain from the probe, mediated by the target protein, leading to the separation of the antisense oligonucleotide from the double-stranded complex. This process triggers the restoration of the fluorescence signal, enabling sensitive detection, and the probe exhibits excellent response within a linear range of GST activity ranging from 0 to 1500 U/L. The results show that not only an efficient strategy for screening robust and practicable aptamers but also an ultrahighly sensitive detection platform for GST was established.

基于色聚体的生物传感器已被广泛构建和应用于检测各种目标。谷胱甘肽 S-转移酶(GST)是一种关键的第二阶段代谢酶,在体内生物转化中起着关键作用,而 GST 的异常表达与各种健康风险有关。在本研究中,我们采用生物素化的 GST 固定化链霉亲和素琼脂糖(SA)珠 SELEX 技术,从一个 79 个核苷酸(nt)的随机单链 DNA(ssDNA)文库中系统地筛选出了靶向 GST 的适配体。经过八轮严格筛选,四种具有惊人相似二级结构的适配体脱颖而出。其中,Seq3 对 GST 的亲和力最高,被选中进行进一步优化。随后,在碱基组成分析、二级结构分析和亲和力评估的基础上,采用了半合理的 SELEX 后截短策略。这一策略在不影响 Seq3 亲和力的前提下,系统地删除了 Seq3 中的冗余核苷酸,最终得到了截短的适配体 Seq3-3,它以 39nt 的紧凑长度保持了特异性。在 Seq3-3 的基础上,我们巧妙地设计了一种双链荧光适配体探针,用于体外检测 GST。其检测机制取决于目标蛋白对探针互补链的竞争性置换,从而导致反义寡核苷酸从双链复合物中分离出来。这一过程会触发荧光信号的恢复,从而实现灵敏的检测,探针在 GST 活性的线性范围(0 至 1500 U/L)内表现出极佳的响应。研究结果表明,该方法不仅为筛选稳健实用的适配体提供了有效策略,而且还建立了一个超高灵敏度的 GST 检测平台。
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引用次数: 0
Advanced Electrochemical Monitoring of Carbendazim Fungicide in Foods Using Interfacial Superassembly of NRPC/NiMn Frameworks. 利用 NRPC/NiMn 框架的界面超组装对食品中的多菌灵杀菌剂进行先进的电化学监测
IF 4.9 3区 工程技术 Q1 CHEMISTRY, ANALYTICAL Pub Date : 2024-10-02 DOI: 10.3390/bios14100474
Shakila Parveen Asrafali, Thirukumaran Periyasamy, Seong Cheol Kim, Jaewoong Lee

A simple, sensitive and reliable sensing system based on nitrogen-rich porous carbon (NRPC) and transition metals, NRPC/Ni, NRPC/Mn and NRPC/NiMn was developed and successfully applied as electrode materials for the quantitative determination of carbendazim (CBZ). The synergistic effect of NRPC and bimetals with acceptable pore structure together with flower-like morphology resulted in producing a highly conductive and interconnected network in NRPC/NiMn@GCE, which significantly enhanced the detection performance of CBZ. The electrochemical behavior investigated by cyclic voltammetry (CV) showed improved CBZ detection for NRPC/NiMn, due to the controlled adsorption/diffusion process of CBZ by the NRPC/NiMn@GCE electrode. The influences of various factors such as pH, NRPC/NiMn concentration, CBZ concentration and scan rate were studied. Under optimal conditions, 0.1 M phosphate-buffered saline (PBS) with a pH of 7.0 containing 30 µg/mL NRPC/NiMn, a favourable linear range detection of CBZ from 5 to 50 µM was obtained. Moreover, a chronoamperometric analysis showed excellent repeatability, reproducibility and anti-interfering ability of the fabricated NRPC/NiMn@GCE sensor. Furthermore, the sensor showed satisfactory results for CBZ detection in real samples with acceptable recoveries of 96.40-104.98% and low RSD values of 0.25-3.45%.

基于富氮多孔碳(NRPC)和过渡金属 NRPC/Ni、NRPC/Mn 和 NRPC/NiMn 开发了一种简单、灵敏和可靠的传感系统,并成功地将其作为电极材料用于多菌灵(CBZ)的定量检测。NRPC 和具有可接受孔隙结构的双金属的协同作用以及花状形貌使 NRPC/NiMn@GCE 中产生了高导电性和互连网络,从而显著提高了 CBZ 的检测性能。循环伏安法(CV)研究的电化学行为表明,由于 NRPC/NiMn@GCE 电极对 CBZ 的吸附/扩散过程进行了控制,NRPC/NiMn@GCE 电极对 CBZ 的检测性能得到了改善。研究了 pH 值、NRPC/NiMn 浓度、CBZ 浓度和扫描速率等各种因素的影响。在 pH 值为 7.0 的 0.1 M 磷酸盐缓冲盐水(PBS)和含有 30 µg/mL NRPC/NiMn 的最佳条件下,CBZ 的线性检测范围为 5 至 50 µM。此外,计时器分析表明,所制造的 NRPC/NiMn@GCE 传感器具有出色的重复性、再现性和抗干扰能力。此外,该传感器在实际样品中的 CBZ 检测结果令人满意,回收率为 96.40-104.98%,RSD 值低至 0.25-3.45%。
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引用次数: 0
A Multi-Channel Urine Sensing Detection System Based on Creatinine, Uric Acid, and pH. 基于肌酸酐、尿酸和 pH 值的多通道尿液传感检测系统
IF 4.9 3区 工程技术 Q1 CHEMISTRY, ANALYTICAL Pub Date : 2024-10-02 DOI: 10.3390/bios14100473
Qiya Gao, Jie Fu, Fangying Xiong, Jiawang Wang, Ziyue Qin, Shuang Li

Urine analysis represents a crucial diagnostic technique employed in clinical laboratories. Creatinine and uric acid in urine are essential biomarkers in the human body and are widely utilized in clinical analysis. Research has demonstrated a correlation between the normal physiological concentrations of creatinine and uric acid in urine and an increased risk of hypertension, cardiovascular diseases, and kidney disease. Furthermore, the pH of urine indicates the body's metabolic processes and homeostatic balance. In this study, an integrated multi-channel electrochemical sensing system was developed, combining electrochemical analysis techniques, microelectronic design, and nanomaterials. The architecture of an intelligent medical detection system and the production of an interactive interface for smartphones were accomplished. Initially, multi-channel selective electrodes were designed for creatinine, uric acid, and pH detection. The detection range was 10 nM to 100 μM for creatinine, 100 μM to 500 μM for uric acid, and 4 to 9 for pH. Furthermore, interference experiments were also conducted to verify the specificity of the sensors. Subsequently, multi-channel double-sided sensing electrodes and function-integrated hardware were designed, with the standard equations of target analytes stored in the system's read-only memory. Moreover, a WeChat mini-program platform was developed for smartphone interaction, enabling off-body detection and real-time display of target analytes through smartphones. Finally, the aforementioned electrochemical detection electrodes were integrated with the smart sensing system and wirelessly interfaced with smartphones, allowing for intelligent real-time detection in primary healthcare and individual household settings.

尿液分析是临床实验室采用的一项重要诊断技术。尿液中的肌酐和尿酸是人体重要的生物标志物,被广泛用于临床分析。研究表明,尿液中肌酐和尿酸的正常生理浓度与罹患高血压、心血管疾病和肾脏疾病的风险增加之间存在相关性。此外,尿液的酸碱度还表明了人体的新陈代谢过程和体内平衡。本研究结合电化学分析技术、微电子设计和纳米材料,开发了一个集成的多通道电化学传感系统。完成了智能医疗检测系统的架构和智能手机交互界面的制作。最初设计的多通道选择性电极用于检测肌酐、尿酸和 pH 值。肌酐的检测范围为 10 nM 至 100 μM,尿酸的检测范围为 100 μM 至 500 μM,pH 值的检测范围为 4 至 9。此外,还进行了干扰实验,以验证传感器的特异性。随后,设计了多通道双面传感电极和功能集成硬件,并在系统的只读存储器中存储了目标分析物的标准方程。此外,还开发了与智能手机交互的微信小程序平台,通过智能手机实现了目标分析物的体外检测和实时显示。最后,将上述电化学检测电极与智能传感系统集成,并与智能手机无线连接,实现了基层医疗机构和个人家庭的智能实时检测。
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