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  • 文章类型: Journal Article
    需要对未经批准的转基因(GM)事件的引入进行监测,因为转基因事件的批准状态可能因国家而异。环介导等温扩增(LAMP)等现场方法为实验室以外的快速GM检测提供了技术答案。较早报道了检测一个GM靶标的实时LAMP测定。为了提高检测的效率,开发了同时靶向花叶病毒启动子(P-FMV)的多重实时LAMP,该启动子构建了花椰菜花叶病毒35S启动子和cry1Ac基因(p35S-cry1Ac)和新霉素磷酸转移酶II(nptII)标记基因之间的区域。该测定可以在45分钟内检测每个GM靶标的低至0.1%。据我们所知,本文首次报道了使用GenieII系统在GM检测中适用性的实时LAMP中的多路复用。所开发的方法提供了快速,现场,以及种子和食品中的实时转基因检测。
    Monitoring of the introduction of unapproved genetically modified (GM) events is required because the approval status of a GM event may differ from country to country. The on-site methods such as loop-mediated isothermal amplification (LAMP) offer a technological answer for the rapid GM detection beyond the laboratories. Real-time LAMP assays detecting one GM target were reported earlier. To increase the efficiency of the assay, a multiplex real-time LAMP simultaneously targeting Figwort Mosaic Virus promoter (P-FMV) that constructs region between the Cauliflower Mosaic Virus 35S promoter and cry1Ac gene (p35S-cry1Ac) and neomycin phosphotransferase II (nptII) marker gene was developed. The assay could detect as low as 0.1% for each GM target within 45 min. To the best of our knowledge, multiplexing in real-time LAMP using the Genie II system with applicability in GM detection has been reported herein for the first time. The developed method provides rapid, on-site, and real-time GM detection in seeds and food products.
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  • 文章类型: Journal Article
    基于等温核酸扩增的侧向流动测试(INAA-LFT)已成为现场病原体检测的强大技术,提供病原体核酸扩增的可见指示,可与实时定量PCR的灵敏度相媲美甚至超越。INAA-LFT的等温性质确保了核酸扩增的一致条件,将其确立为快速现场病原体检测的关键技术。然而,尽管有相当大的希望,基于INAA扩增的等温横向流动测试的广泛应用面临着几个挑战。本综述概述了INAA-LFT程序,强调其在检测植物病毒方面的进步。此外,审查强调了解决现有局限性的必要性,并强调了正在进行的研究工作,致力于提高该技术在快速现场测试领域的适用性和性能。
    Isothermal nucleic acid amplification-based lateral flow testing (INAA-LFT) has emerged as a robust technique for on-site pathogen detection, providing a visible indication of pathogen nucleic acid amplification that rivals or even surpasses the sensitivity of real-time quantitative PCR. The isothermal nature of INAA-LFT ensures consistent conditions for nucleic acid amplification, establishing it as a crucial technology for rapid on-site pathogen detection. However, despite its considerable promise, the widespread application of isothermal INAA amplification-based lateral flow testing faces several challenges. This review provides an overview of the INAA-LFT procedure, highlighting its advancements in detecting plant viruses. Moreover, the review underscores the imperative of addressing the existing limitations and emphasizes ongoing research efforts dedicated to enhancing the applicability and performance of this technology in the realm of rapid on-site testing.
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  • 文章类型: Journal Article
    由于Covid-19病毒的快速传播,为了避免疾病的传播,教育系统经历了许多变化。教育机构已经从传统的教学教学法转向在线学习过程。这项研究的目的是评估在线学习数学和高等教育现场学习数学之间的差异。要确定在线和现场学习数学之间是否有区别,我们首先分析了完成同一数学课程的学生的成绩。然后我们分析了成功完成本课程的学生的成绩。学生的学习成绩是由数学课程考试的最终成绩来衡量的,由科索沃UBT大学计算机科学与工程系提供,这所大学的学生是这项研究的一部分。这项研究为解决在线和现场数学学习与成就之间的差异问题做出了重要贡献。调查结果显示,在线学习成果和现场学习成果之间存在微小差异。结果表明,在线学习者在算术课程中的表现优于现场学习者。这项研究的结果将为希望提供满足学生和社区要求的一流在线课程的高等教育机构提供重要信息。
    As a result of the rapid spread of the Covid-19 virus, the education system has undergone many changes in order to avoid the spread of the disease. Educational institutions have moved from traditional pedagogy of teaching to the process of online learning. The purpose of this research is to estimate there are differences between learning mathematics online and on-site in Higher Education. To determine if there is a difference between learning math online and on-site, we first analyzed the results of students who have completed the same math course. Then we analyzed the results of students who have successfully completed this course. The students\' academic results were measured by the final grade of the exams from the mathematics course, which were provided by the Department of Computer Science and Engineering at UBT University in Kosovo, where the students of this University were part of this study. This research study makes an important contribution to addressing the question of differences between online and on-site mathematics learning and achievement. The findings revealed that there are small differences between online learning outcomes and on-site. The outcomes demonstrated that online learners performed better than on-site learners in the arithmetic course. The results of this study will offer crucial information to higher education institutions looking to provide top-notch online courses that cater to students\' and communities\' requirements.
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  • 文章类型: Journal Article
    智能化的发展,敏感,和视觉方法快速检测兽药残留对于确保食品质量和安全至关重要。这里,提出了一种基于智能手机的双反信号MOFs荧光传感系统,用于孔雀石绿(MG)的智能现场视觉检测。采用简单的一锅法一步成功合成了UiO-66-NH2@RhB-双发射荧光探针。内部过滤效应(IFE)猝灭红色荧光,而氢键相互作用增强了蓝色荧光,实现高度敏感,准确,并通过荧光分析直观检测MG双反信号。探针在0.1-100μmol/L的宽范围内显示出良好的线性,检测限(LOD)为20nmol/L通过集成智能手机摄影和RGB(红色,绿色,和蓝色)分析,可以在5分钟内实现对水中和实际鱼样中MG的准确定量分析。这个开发的平台为MG在实际应用中的现场检测提供了巨大的希望,具有简单的优点,成本效益,和速度。因此,它可能为现场评估食品安全和环境健康开辟一条新途径。
    The development of intelligent, sensitive, and visual methods for the rapid detection of veterinary drug residues is essential to ensure food quality and safety. Here, a smartphone-based dual inverse signal MOFs fluorescence sensing system was proposed for intelligent in-site visual detection of malachite green (MG). A UiO-66-NH2@RhB-dual-emission fluorescent probe was successfully synthesized in one step using a simple one-pot method. The inner filter effect (IFE) quenches the red fluorescence, while hydrogen bonding interaction enhances the blue fluorescence, enabling highly sensitive, accurate, and visual detection of MG dual inverse signals through fluorescence analysis. The probe showed great linearity over a wide range of 0.1-100 μmol/L, with a limit of detection (LOD) of 20 nmol/L. By integrating smartphone photography and RGB (red, green, and blue) analysis, accurate quantitative analysis of MG in water and actual fish samples can be achieved within 5 min. This developed platform holds great promise for the on-site detection of MG in practical applications, with the advantages of simplicity, cost-effectiveness, and rapidity. Consequently, it may open up a new pathway for on-site evaluation of food safety and environmental health.
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  • 文章类型: Journal Article
    全球数百万人受到地下水中天然存在的砷的影响。开发一种低成本、高度敏感,用于快速现场检测水中砷的便携式测定法对于确定安全井的区域并帮助确定测试的优先级很重要。在这里,开发了一种新型的基于纸张的荧光测定法,用于砷的现场分析,它是由AsH3的固相荧光滤光片效应(SPFFE)构建的,它诱导了向碳点的银纳米颗粒(AgNPs)的生成。由于AsH3对Ag的有效还原和AgNP的高摩尔消光系数,拟议的基于SPFFE的测定实现了0.36μg/L的低砷检测限。结合智能手机和集成的样品处理和传感平台,可以实现对砷的现场敏感检测。通过成功分析地表和地下水样品,验证了便携式测定的准确性,与通过质谱获得的结果没有显着差异。与其他砷分析方法相比,这个开发的系统提供了出色的灵敏度,便携性,和低成本。它具有对地下水中砷进行现场分析的潜力,可以确定安全的井位,并从全球地下水砷地图中快速获得产出。
    Millions of people worldwide are affected by naturally occurring arsenic in groundwater. The development of a low-cost, highly sensitive, portable assay for rapid field detection of arsenic in water is important to identify areas for safe wells and to help prioritize testing. Herein, a novel paper-based fluorescence assay was developed for the on-site analysis of arsenic, which was constructed by the solid-phase fluorescence filter effect (SPFFE) of AsH3-induced the generation of silver nanoparticles (AgNPs) toward carbon dots. The proposed SPFFE-based assay achieves a low arsenic detection limit of 0.36 μg/L due to the efficient reduction of Ag+ by AsH3 and the high molar extinction coefficient of AgNPs. In conjunction with a smartphone and an integrated sample processing and sensing platform, field-sensitive detection of arsenic could be achieved. The accuracy of the portable assay was validated by successfully analyzing surface and groundwater samples, with no significant difference from the results obtained through mass spectrometry. Compared to other methods for arsenic analysis, this developed system offers excellent sensitivity, portability, and low cost. It holds promising potential for on-site analysis of arsenic in groundwater to identify safe well locations and quickly obtain output from the global map of groundwater arsenic.
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  • 文章类型: Journal Article
    及时的万古霉素治疗药物监测(TDM)可以迅速调整剂量和安全治疗。当地事件促使人们调查报告时间延长的原因。
    为了调查有和没有现场TDM处理的医院之间万古霉素浓度报告时间的变化,和患者安全问题。
    医院1的万古霉素浓度结果(非现场监测),对2021年6月至12月的2号和3号医院(均为现场监测)进行了回顾性分析。在现场TDM开始后3个月对医院重复回顾性数据收集以进行比较。对医院1的万古霉素临床事件进行了审查,以确定何时延迟报告结果可能导致患者不良结局的例子。
    与医院2和医院T3相比,医院1的中位报告时间为11.13h(分别为1.73h和1.70h)。在医院1开始现场TDM后,报告时间减少至1.33h(p<0.001)。在非现场监测期间,第一医院发生的几起事件涉及TDM结果的延迟。
    TDM的非现场处理导致万古霉素浓度报告明显延迟,通过过渡到现场测试的可用性,这一点得到了显着改善。这项研究还强调了准确的问题识别对提高患者安全性的影响。
    UNASSIGNED: Timely vancomycin therapeutic drug monitoring (TDM) enables prompt dose adjustments and safe treatment. Local incidents prompted an investigation into the reasons for prolonged reporting times.
    UNASSIGNED: To investigate the variation in reporting times of vancomycin concentrations between hospitals with and without on-site TDM processing, and patient safety implications.
    UNASSIGNED: Vancomycin concentration results for Hospital 1 (off-site monitoring), Hospitals 2 and 3 (both on-site monitoring) from June to December 2021 were retrospectively analysed. Retrospective data collection was repeated for Hospital 1 three months post on-site TDM commencement for comparison. Vancomycin clinical incidents at Hospital 1 were reviewed to identify examples of when delays in reporting of results potentially contributed towards adverse patient outcomes.
    UNASSIGNED: Hospital 1 had a median reporting time of 11.13 h compared with Hospital 2 and Hospital T3 (1.73 h and 1.70 h respectively). Following the commencement of on-site TDM at Hospital 1, the reporting time reduced to 1.33 h (p < 0.001). Several incidents at Hospital 1 during the period of off-site monitoring involved delays to TDM results.
    UNASSIGNED: Off-site processing of TDM introduced significant delays in reporting of vancomycin concentrations, which was significantly improved by transitioning to onsite availability of testing. This study also highlights the impact of accurate problem identification in improving patient safety.
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  • 文章类型: Journal Article
    木材防腐剂的定性和定量现场检测方法对回收行业和职业健康安全具有很高的价值。经过数十年的使用,木材防腐剂对人类和环境有毒。为了检测受污染的木材以及将成熟的木材加工成刨花板,需要一种通用的检测方法。特别是历史上的木制品用防腐剂如五氯苯酚处理,林丹或二氯二苯基三氯乙烷。这就需要一种非破坏性的现场检测方法,这不需要专门的人员。在该出版物中,提出了两种通过固相微萃取利用顶空采样的方法,随后使用气相色谱进行分离,并通过漂移管离子迁移谱仪(SPME-HS-GC-DTIMS)进行检测。一种方法能够定量检测木粉和木屑中的五氯苯酚,因为它们用于木材加工行业。使用DIN32645实现0.1mg/kg的检测限,其可以甚至更低。第二种方法可以实现五氯苯酚的无损检测,林丹,木质物体中的二氯二苯基三氯乙烷和其他防腐剂。因此,准备好样品,其浓度明显低于典型的治疗对象,并在实际样品旁边用于方法验证。证明了该方法对实际样品和低浓度制备样品的污染。
    Qualitative and quantitative on-site detection methods for wood preservatives are of high value for the recycling industry and the occupational health and safety. Wood preservatives revealed as toxic to human and environment after decades of use. For the detection of contaminated wood and for processing of matured timber to particle boards a versatile detection method is needed. Especially historical wooden objects were treated with preservatives like pentachlorophenol, lindane or dichlorodiphenyltrichloroethane. This requires a non-destructive on-site detection method, that does not require specialized personnel. In this publication two methods are presented utilizing headspace sampling by solid-phase microextraction, subsequent separation using gas chromatography and detection by a drift tube ion mobility spectrometer (SPME-HS-GC-DTIMS). One method enables the quantitative detection of pentachlorophenol in wood flour and wood chips as they are used in wood processing industries. A limit of detection of 0.1 mg/kg was achieved using DIN 32645, which can be even more lowered. The second method enables non-destructive detection of pentachlorophenol, lindane, dichlorodiphenyltrichloroethane and other preservatives in wooden objects. Therefore, samples were prepared, which show a significantly lower concentration than typical treated objects, and used next to real samples for method validation. With the method contamination of the real samples and of the prepared samples of low concentration were proven.
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  • 文章类型: Journal Article
    考虑到重金属离子(HMIs)通过食物链转移并在牛奶中积累的高风险,一个灵活和方便的即时测试(POCT)平台是迫切需要准确的,敏感,以及牛奶中多种HMI的高度选择性现场定量。在这项工作中,设计了具有六个丝网印刷电极(SPE)的具有成本效益的磁盘,用于手持式电化学检测。采用金属有机骨架(MOFs)放大和增强亚甲基蓝(MB)的电化学信号。使用差分脉冲伏安法(DPV)方法,四种HMI的低检测限(Cd2+,0.039ppb;Hg2+,0.039ppb;Pb2+,0.073ppb;和As3+,0.022ppb)在四分钟内实现。此外,将定量POCT系统应用于牛奶样品。成本低的优点,易于现场实施,快速反应,和准确性允许POCT平台用于牛奶样品中多种HMI的定量的实际监测应用。
    Considering the high risk of heavy metal ions (HMIs) transferring through the food chain and accumulating in milk, a flexible and facile point-of-care testing (POCT) platform is urgently needed for the accurate, sensitive, and highly selective on-site quantification of multiple HMIs in milk. In this work, a cost-effective disk with six screen-printed electrodes (SPEs) was designed for hand-held electrochemical detection. Metal organic frameworks (MOFs) were adopted to amplify and enhance the electrochemical signals of methylene blue (MB). Using differential pulse voltammetry (DPV) methods, low limits of detection for four HMIs (Cd2+, 0.039 ppb; Hg2+, 0.039 ppb; Pb2+, 0.073 ppb; and As3+, 0.022 ppb) were achieved within four minutes. Moreover, the quantitative POCT system was applied to milk samples. The advantages of low cost, ease of on-site implementation, fast response, and accuracy allow for the POCT platform to be used in practical monitoring applications for the quantitation of multiple HMIs in milk samples.
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  • 文章类型: Journal Article
    溶解的硫化物在离开基质时倾向于物种转化和损失,因此,开发实用的现场硫化物测定对环境监测和人类健康至关重要。在这项工作中,开发了一种新型的纸质比率荧光传感器,用于硫化物的现场分析,该系统是通过CdS量子点(QD)对碳点(C点)的内部滤波效应(IFE)构建的。而不是水相系统,硫化物转化为氢化物会诱导CdS量子点在纸上的原位形成,充当能量受体来淬灭C点的发射,随着硫化物浓度的增加,导致比率荧光从蓝色到黄色的变化。此外,我们提出了一种基于智能手机的荧光捕获设备,集成了一个编程的Python程序,实现了硫化物的颜色识别和准确检测。在最优条件下,这种比率荧光传感器可以对硫化物进行现场分析,检出限为0.05μM。通过对环境水样的成功现场分析,以令人满意的回收率验证了传感器的准确性。与其他用于硫化物分析的荧光法相比,这个开发的系统保留了无标签的优点,低成本,操作方便,和小型化,显示出现场测量硫化物的巨大潜力,以及环境监测。
    Dissolved sulfide tends to species transformation and loss upon leaving the matrix, thus the development of a practical on-site determination of sulfide is crucial for environmental monitoring and human health. In this work, a novel paper-based ratiometric fluorescence sensor was developed for the field analysis of sulfide, which system was constructed by the inner filter effect (IFE) of CdS quantum dots (QDs) toward carbon dots (C-dots). Instead of an aqueous phase system, the conversion of sulfide to its hydride would induce the in-situ formation of CdS QDs on the paper, which acted as an energy acceptor to quench the emission of C-dots, leading to a variation of ratiometric fluorescence from blue to yellow with the increasing concentration of sulfide. Moreover, we proposed a smartphone-based fluorescence capture device integrated with a programmed Python program, accomplishing both color recognition and accurate detection of sulfide. Under the optimal condition, this ratiometric fluorescence sensor allowed for the on-site analysis of sulfide with a limit of detection of 0.05 μM. The accuracy of the sensor was validated via the successful field analysis of environmental water samples with satisfactory recoveries. Compared to other fluorescence methods used for sulfide analysis, this developed system retains the advantages of label-free, low-cost, ease of operation, and miniaturization, showing great potential for the measurement of sulfide on-site, as well as environmental monitoring.
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  • 文章类型: Journal Article
    大肠杆菌(E.coli)O157:H7是威胁人类健康的主要食源性和水性致病菌。由于其在低浓度下的高毒性,建立省时、高灵敏度的原位检测方法至关重要。在这里,我们开发了一种快速的,超灵敏,以及基于重组酶辅助扩增(RAA)和CRISPR/Cas12a技术的组合检测大肠杆菌O157:H7的可视化方法。使用RAA方法对基于CRISPR/Cas12a的系统进行预扩增,该方法具有较高的灵敏度,能够检测到低至〜1CFU/mL(荧光法)和1×102CFU/mL(侧流测定)的大肠杆菌O157:H7,远低于传统的检出限。实时PCR技术(103CFU/mL)和ELISA(104〜107CFU/mL)。此外,通过模拟真实牛奶和饮用水样品的检测,证明了该方法在实际样品中仍然具有良好的适用性。重要的是,我们的RAA-CRISPR/Cas12a检测系统可以完成整个过程(包括提取、扩增,和检测)在优化条件下55分钟内,比大多数其他报道的传感器都快,这需要几个小时到几天。信号读出也可以通过用手持式UV灯或肉眼检测的侧流测定产生的荧光可视化,这取决于所使用的DNA报道分子。因为速度快的优点,具有高灵敏度,不需要复杂的设备,该方法对痕量病原体的原位检测具有广阔的应用前景。
    Escherichia coli (E. coli) O157:H7 is a major foodborne and waterborne pathogen that can threaten human health. Due to its high toxicity at low concentrations, it is crucial to establish a time-saving and highly sensitive in situ detection method. Herein, we developed a rapid, ultrasensitive, and visualized method for detecting E. coli O157:H7 based on a combination of Recombinase-Aided Amplification (RAA) and CRISPR/Cas12a technology. The CRISPR/Cas12a-based system was pre-amplified using the RAA method, which showed high sensitivity and enabled detecting as low as ~1 CFU/mL (fluorescence method) and 1 × 102 CFU/mL (lateral flow assay) of E. coli O157:H7, which was much lower than the detection limit of the traditional real-time PCR technology (103 CFU/mL) and ELISA (104~107 CFU/mL). In addition, we demonstrated that this method still has good applicability in practical samples by simulating the detection in real milk and drinking water samples. Importantly, our RAA-CRISPR/Cas12a detection system could complete the overall process (including extraction, amplification, and detection) within 55 min under optimized conditions, which is faster than most other reported sensors, which take several hours to several days. The signal readout could also be visualized by fluorescence generated with a handheld UV lamp or a naked-eye-detected lateral flow assay depending on the DNA reporters used. Because of the advantages of being fast, having high sensitivity, and not requiring sophisticated equipment, this method has a promising application prospect for in situ detection of trace amounts of pathogens.
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