4种植物源性成分多重real-time PCR检测方法的建立及其在食用淀粉中的应用

建立一种可同时快速检测红薯、木薯、马铃薯、玉米源性成分的多重实时聚合酶链式反应(real-time polymerase chain reaction,real-time PCR)方法。分别以红薯g3pdh基因、木薯g3pdh基因、马铃薯UGPase基因、玉米zSSIIb基因为靶基因设计特异性引物和TaqMan探针,以18S rRNA基因为内参基因,建立多重real-time PCR方法,开展方法学验证,并对不同掺入比例模拟样品和实际淀粉样品进行检测。结果显示,该方法具有高通量、特异性强、灵敏度高等优点。与15种非目标源性均无交叉反应;对目标DNA的检测灵敏度可达到3×10^(-3) ng/μL,且具有良好的线性关系和扩增效率;对淀粉样品的检出限可达0.1%,对50份实际样品进行检测,结果与参比方法一致,说明建立的多重real-time PCR法可用于食用淀粉种类掺假鉴别检测。

垂体后叶粉中3种动物源性成分多重real-time PCR检测方法的建立及应用

目的基于三重实时聚合酶链式反应(real-time polymerase chain reaction,real-time PCR)技术同时快速检测垂体后叶粉中的猪、牛、羊性动物源性成分。方法采用CTAB法提取样品及对照品DNA,以线粒体基因为靶基因设计并合成三重real-time PCR的引物探针,并对建立方法进行验证,用于检测垂体后叶粉样品是否与标识一致。结果猪、牛、羊源性成分的最低检测浓度为10 pg/μL,采用建立的方法检测3批垂体后叶粉样品,均只检测出猪源性成分。结论建立的方法可用于同时检测垂体后叶粉中的猪、牛、羊源性成分,可为相关工作提供技术支持。

多重real-time PCR技术快速鉴别特种乳中的乳源动物成分

建立一种特种乳中乳源动物成分快速鉴别多重实时聚合酶链式反应技术。通过筛选建立8种不同乳源物种检测的多重实时聚合酶链式反应方法,在一个反应体系里可同时检测4种乳源的特异性靶基因,绝对灵敏度达0.1~5 pg/μL,检出限可达0.1%。同时,建立了高效的DNA提取方法,样品裂解后可在12 min左右完成96个样品的DNA提取,大大缩短了样品前处理时间。采用该方法对市售特色乳产品进行调查,结果显示标识不准确的产品比例达36.36%。

4种动物源性成分多重real-time PCR检测方法的建立及其在驴肉制品检测中的应用

建立一种同时快速检测驴、马、猪及鸭源性成分的四重实时聚合酶链式反应(real-time polymerase chain reaction,real-time PCR)方法。分别以4种源性成分的Nad5、ATpase6、ATP8、cytb基因为靶基因设计特异性引物和TaqMan探针,以18S rRNA基因为内参基因,建立多重real-time PCR方法,并对该方法进行方法学验证,同时对不同掺入比例模拟样品、不同加工工艺模拟样品和实际驴肉样品进行检测。结果显示,该方法具有高通量、特异性强、灵敏度高等优点。当Ct值≤35.0时,方法对16种非目标源性具有良好特异性;灵敏度可检测到质量浓度为2×10^(-4)ng/μL的模板DNA;对生肉的检出限为肉含量的0.001%,对熟肉制品的检出限为肉含量的0.01%;对100份实际样品进行检测,结果与标准方法一致,说明建立的多重real-time PCR法可用于肉及肉制品中常见掺假源性成分的检测。

3种致病菌多重real-time PCR检测方法的建立及其在散装即食肉制品中的应用

建立一种同时快速检测沙门氏菌、金黄色葡萄球菌和蜡样芽孢杆菌的三重实时聚合酶链式反应(realtime polymerase chain reaction,real-time PCR)方法。以沙门氏菌invA基因、金黄色葡萄球菌Sa442基因和蜡样芽孢杆菌Cereolysin AB基因为靶基因设计引物和TaqMan探针,建立多重real-time PCR方法,对该方法进行方法学验证,并对实际的散装即食肉制品样品进行检测。结果表明,该方法特异性强、灵敏度高、重复性好。对15株非目标菌进行检测,结果均为阴性;3种致病菌的定量线性浓度范围为10^(2)~10^(8) CFU/mL,且定量检测批内和批间的变异系数均小于2%;沙门氏菌、金黄色葡萄球菌和蜡样芽孢杆菌在未增菌的即食肉制品样品中检出限分别为3.8×10^(2)、4.9×10^(2) CFU/mL和5.7×10^(2) CFU/mL,经增菌5 h后,检出限提高到3.8、4.9 CFU/mL和5.7 CFU/mL。对100份实际样品进行检测,结果与标准方法一致,说明建立的多重real-time PCR法可以用于散装即食肉制品中3种致病菌的检测。

应用多重荧光Real-time PCR快速鉴定猪链球菌2型

目的建立能同时鉴定链球菌2型(S.suis2)和确定主要毒力标志MRP的快速检测方法。方法根据S.suis2CPS2J与MRP的保守序列,设计并合成了2对引物及其相应探针。通过优化各反应物的浓度、配比和循环程序,建立能从组织中直接检测CPS2J和MRP的实时荧光PCR方法。结果能在3h内对送检组织样品检测确定S.suis2型和MRP毒力基因,最低检测菌体浓度24CFU/ml。与普通PCR相比,敏感性在检测CPS2J时比普通PCR高至少10倍以上,而在检测MRP时比其高100倍。与葡萄球菌、化脓链球菌、沙门氏菌、S.suis1型、9型等无交叉反应。同一人在试剂放置0d、20d和60d检测,以及不同人用同一方法检测,变异系数均小于5%,差异不显著。用于检测SSsc0501攻毒后的样品,结果在心、肝、脾、肺、肾、淋巴结、脑中均检测到S.suis2阳性,MRP阳性。而在肌肉中,S.suis2和MRP均为阴性。结论建立的多重荧光Real-timePCR敏感性高、特异性强,稳定性和重复性好,能用于可疑S.suis2型组织样品的快速定型和毒力鉴定。

基于Real-time PCR法检测乳粉中牛源性成分定量研究

基于Real-timePCR建立了乳粉中牛源性成分相对定量检测方法,并对牛的特异性引物与探针进行了特异性、灵敏度和稳定性测试。通过模拟不同浓度牛乳粉与马乳粉混合样本,根据其△Ct值的函数关系进行线性拟合进而绘制标准曲线,建立乳粉中牛源性成分的相对定量检测。结果显示,该方法的最低检测限为0.00001 mg/mL,回收率为91.11%~119.2%,组间变异系数≤0.58%、组内变异系数≤1.44%。说明该方法在特异性与稳定性上适用于乳粉中牛源性成分及含量的掺假检测。

A multifunctional shear apparatus for rocks subjected to true triaxial stress and high temperature in real-time

Deep engineering disasters,such as rockbursts and collapses,are more related to the shear slip of rock joints.A novel multifunctional device was developed to study the shear failure mechanism in rocks.Using this device,the complete shearedeformation process and long-term shear creep tests could be performed on rocks under constant normal stiffness(CNS)or constant normal loading(CNL)conditions in real-time at high temperature and true-triaxial stress.During the research and development process,five key technologies were successfully broken through:(1)the ability to perform true-triaxial compressioneshear loading tests on rock samples with high stiffness;(2)a shear box with ultra-low friction throughout the entire stress space of the rock sample during loading;(3)a control system capable of maintaining high stress for a long time and responding rapidly to the brittle fracture of a rock sample as well;(4)a refined ability to measure the volumetric deformation of rock samples subjected to true triaxial shearing;and(5)a heating system capable of maintaining uniform heating of the rock sample over a long time.By developing these technologies,loading under high true triaxial stress conditions was realized.The apparatus has a maximum normal stiffness of 1000 GPa/m and a maximum operating temperature of 300C.The differences in the surface temperature of the sample are constant to within5C.Five types of true triaxial shear tests were conducted on homogeneous sandstone to verify that the apparatus has good performance and reliability.The results show that temperature,lateral stress,normal stress and time influence the shear deformation,failure mode and strength of the sandstone.The novel apparatus can be reliably used to conduct true-triaxial shear tests on rocks subjected to high temperatures and stress.

Pore-pressure and stress-coupled creep behavior in deep coal:Insights from real-time NMR analysis

Understanding the variations in microscopic pore-fracture structures(MPFS) during coal creep under pore pressure and stress coupling is crucial for coal mining and effective gas treatment. In this manuscript, a triaxial creep test on deep coal at various pore pressures using a test system that combines in-situ mechanical loading with real-time nuclear magnetic resonance(NMR) detection was conducted.Full-scale quantitative characterization, online real-time detection, and visualization of MPFS during coal creep influenced by pore pressure and stress coupling were performed using NMR and NMR imaging(NMRI) techniques. The results revealed that seepage pores and microfractures(SPM) undergo the most significant changes during coal creep, with creep failure gradually expanding from dense primary pore fractures. Pore pressure presence promotes MPFS development primarily by inhibiting SPM compression and encouraging adsorption pores(AP) to evolve into SPM. Coal enters the accelerated creep stage earlier at lower stress levels, resulting in more pronounced creep deformation. The connection between the micro and macro values was established, demonstrating that increased porosity at different pore pressures leads to a negative exponential decay of the viscosity coefficient. The Newton dashpot in the ideal viscoplastic body and the Burgers model was improved using NMR experimental results, and a creep model that considers pore pressure and stress coupling using variable-order fractional operators was developed. The model’s reasonableness was confirmed using creep experimental data. The damagestate adjustment factors ω and β were identified through a parameter sensitivity analysis to characterize the effect of pore pressure and stress coupling on the creep damage characteristics(size and degree of difficulty) of coal.

一种基于real-time PCR技术的TTV检测方法的建立及应用

目的:本研究旨在开发一种具有更高灵敏度和特异性的TTV检测技术,为揭示TTV在多种疾病过程中的作用提供重要的技术支持。方法:为了更精确、灵敏的检测TTV,本研究分析了目前公布的所有亚型的TTV基因序列,在此基础上建立了一种基于UTR区域的real-time PCR检测方法,并与文献报道应用较为广泛的PCR检测方法进行了对比。结果:本研究建立的方法在1×10^(7)~1×10^(1) copies/μL标准品浓度范围内具有良好的线性关系,相关系数为1.000,斜率为-3.446,检测下限为1×10^(1) copies/μL。重复性试验结果显示,组内变异系数为7.22%,表明本方法重复性、稳定性较强。针对30份临床样本,使用本研究建立的real-time PCR检测方法及目前被多个研究所使用的4套引物进行对比。结果表明,本研究所建立的方法灵敏度显著高于文献中报道的4种方法(P<0.01);Sanger测序结果表明,本方法检测出的30份阳性样本均为TTV,检测特异性为100%。结论:本研究采用基于TaqMan探针的real-time PCR检测方法,检测灵敏性高、覆盖基因型范围广,尤其对于TTV病毒载量较低的情况下能够进行定量检测,对于TTV病毒的致病性及作为免疫标志物的应用提供重要的技术支持。

Real-time data processing method for CO_(2) dispersion interferometer on EAST

A real-time data processing system is designed for the carbon dioxide dispersion interferometer(CO_(2)-DI)on EAST.The system utilizes the parallel and pipelining capabilities of an fieldprogrammable gate array(FPGA)to digitize and process the intensity of signals from the detector.Finally,the real-time electron density signals are exported through a digital-to-analog converter(DAC)module in the form of analog signals.The system has been successfully applied in the CO_(2)-DI system to provide low-latency electron density input to the plasma control system on EAST.Experimental results of the latest campaign with long-pulse discharges on EAST(2022–2023)demonstrate that the system can respond effectively in the case of rapid density changes,proving its reliability and accuracy for future electron density calculation.

Real-Time Intelligent Diagnosis of Co-frequency Vibration Faults in Rotating Machinery Based on Lightweight-Convolutional Neural Networks

The co-frequency vibration fault is one of the common faults in the operation of rotating equipment,and realizing the real-time diagnosis of the co-frequency vibration fault is of great significance for monitoring the health state and carrying out vibration suppression of the equipment.In engineering scenarios,co-frequency vibration faults are highlighted by rotational frequency and are difficult to identify,and existing intelligent methods require more hardware conditions and are exclusively time-consuming.Therefore,Lightweight-convolutional neural networks(LW-CNN)algorithm is proposed in this paper to achieve real-time fault diagnosis.The critical parameters are discussed and verified by simulated and experimental signals for the sliding window data augmentation method.Based on LW-CNN and data augmentation,the real-time intelligent diagnosis of co-frequency is realized.Moreover,a real-time detection method of fault diagnosis algorithm is proposed for data acquisition to fault diagnosis.It is verified by experiments that the LW-CNN and sliding window methods are used with high accuracy and real-time performance.

Nanomaterial-assisted wearable glucose biosensors for noninvasive real-time monitoring:Pioneering point-of-care and beyond

This review explores glucose monitoring and management strategies,emphasizing the need for reliable and userfriendly wearable sensors that are the next generation of sensors for continuous glucose detection.In addition,examines key strategies for designing glucose sensors that are multi-functional,reliable,and cost-effective in a variety of contexts.The unique features of effective diabetes management technology are highlighted,with a focus on using nano/biosensor devices that can quickly and accurately detect glucose levels in the blood,improving patient treatment and control of potential diabetes-related infections.The potential of next-generation wearable and touch-sensitive nano biomedical sensor engineering designs for providing full control in assessing implantable,continuous glucose monitoring is also explored.The challenges of standardizing drug or insulin delivery doses,low-cost,real-time detection of increased blood sugar levels in diabetics,and early digital health awareness controls for the adverse effects of injectable medication are identified as unmet needs.Also,the market for biosensors is expected to expand significantly due to the rising need for portable diagnostic equipment and an ever-increasing diabetic population.The paper concludes by emphasizing the need for further research and development of glucose biosensors to meet the stringent requirements for sensitivity and specificity imposed by clinical diagnostics while being cost-effective,stable,and durable.

Integrated strategy for real-time wind power fluctuation mitigation and energy storage system control

To address the impact of wind-power fluctuations on the stability of power systems,we propose a comprehensive approach that integrates multiple strategies and methods to enhance the efficiency and reliability of a system.First,we employ a strategy that restricts long-and short-term power output deviations to smoothen wind power fluctuations in real time.Second,we adopt the sliding window instantaneous complete ensemble empirical mode decomposition with adaptive noise(SW-ICEEMDAN)strategy to achieve real-time decomposition of the energy storage power,facilitating internal power distribution within the hybrid energy storage system.Finally,we introduce a rule-based multi-fuzzy control strategy for the secondary adjustment of the initial power allocation commands for different energy storage components.Through simulation validation,we demonstrate that the proposed comprehensive control strategy can smoothen wind power fluctuations in real time and decompose energy storage power.Compared with traditional empirical mode decomposition(EMD),ensemble empirical mode decomposition(EEMD),and complete ensemble empirical mode decomposition with adaptive noise(CEEMDAN)decomposition strategies,the configuration of the energy storage system under the SW-ICEEMDAN control strategy is more optimal.Additionally,the state-of-charge of energy storage components fluctuates within a reasonable range,enhancing the stability of the power system and ensuring the secure operation of the energy storage system.

Real-time debris flow monitoring and automated warning system

At present,debris flow warning uses precipitation threshold and issues regional warning throughout the world.Precipitation threshold warning is less accurate and in most of the time large portion of unaffected population are evacuated.More precise warning should use direct monitoring.There are many debris flow monitoring stations but no real time warning system in use.The main reason is that the identification and confirmation of debris flow occurrence requires human interaction and it is too slow.A debris flow monitoring and warning system has been installed in the midstream section of Yusui Stream,Taiwan China.The monitoring station operates fully automatically,providing early warnings without the need for manual intervention.The system comprises two webcam cameras,two Micro-Electro-Mechanical Systems(MEMS),and a rain gauge.The arrival of debris flows is detected and confirmed through both webcam images and MEMS signals.Once debris flow is detected,the system automatically issues a warning to the affected areas via voice messages,line messages,broadcasts,and web-based alerts.The webcam cameras are also used to estimate debris flow velocity and flow height,while the MEMS sensors are utilized to determine the phase speed and flow rate.On July 24th,2014,Typhoon Gaemi triggered several debris flows,and the system successfully issued several warnings automatically.The entire video record,along with depth variation data,was recorded automatically.

Real-Time Detection and Instance Segmentation of Strawberry in Unstructured Environment

The real-time detection and instance segmentation of strawberries constitute fundamental components in the development of strawberry harvesting robots.Real-time identification of strawberries in an unstructured envi-ronment is a challenging task.Current instance segmentation algorithms for strawberries suffer from issues such as poor real-time performance and low accuracy.To this end,the present study proposes an Efficient YOLACT(E-YOLACT)algorithm for strawberry detection and segmentation based on the YOLACT framework.The key enhancements of the E-YOLACT encompass the development of a lightweight attention mechanism,pyramid squeeze shuffle attention(PSSA),for efficient feature extraction.Additionally,an attention-guided context-feature pyramid network(AC-FPN)is employed instead of FPN to optimize the architecture’s performance.Furthermore,a feature-enhanced model(FEM)is introduced to enhance the prediction head’s capabilities,while efficient fast non-maximum suppression(EF-NMS)is devised to improve non-maximum suppression.The experimental results demonstrate that the E-YOLACT achieves a Box-mAP and Mask-mAP of 77.9 and 76.6,respectively,on the custom dataset.Moreover,it exhibits an impressive category accuracy of 93.5%.Notably,the E-YOLACT also demonstrates a remarkable real-time detection capability with a speed of 34.8 FPS.The method proposed in this article presents an efficient approach for the vision system of a strawberry-picking robot.

A real-time performance improvement method for composite time scale

The composite time scale(CTS)provides a stable,accurate,and reliable time scale for modern society.The improvement of CTS’s real-time performance will improve its stability,which strengths related applications’performance.Aiming at this goal,a method achieved by determining the optimal calculation interval and accelerating adjustment stage is proposed in this paper.The determinants of the CTS’s calculation interval(characteristics of the clock ensemble,the measurement noise,the time and frequency synchronization system’s noise and the auxiliary output generator noise floor)are studied and the optimal calculation interval is obtained.We also investigate the effect of ensemble algorithm’s initial parameters on the CTS’s adjustment stage.A strategy to get the reasonable initial parameters of ensemble algorithm is designed.The results show that the adjustment stage can be finished rapidly or even can be shorten to zero with reasonable initial parameters.On this basis,we experimentally generate a distributed CTS with a calculation interval of 500 s and its stability outperforms those of the member clocks when the averaging time is longer than1700 s.The experimental result proves that the CTS’s real-time performance is significantly improved.

Models to Simulate Effective Coverage of Fire Station Based on Real-Time Travel Times

In recent years,frequent fire disasters have led to enormous damage in China.Effective firefighting rescues can minimize the losses caused by fires.During the rescue processes,the travel time of fire trucks can be severely affected by traffic conditions,changing the effective coverage of fire stations.However,it is still challenging to determine the effective coverage of fire stations considering dynamic traffic conditions.This paper addresses this issue by combining the traveling time calculationmodelwith the effective coverage simulationmodel.In addition,it proposes a new index of total effective coverage area(TECA)based on the time-weighted average of the effective coverage area(ECA)to evaluate the urban fire services.It also selects China as the case study to validate the feasibility of the models,a fire station(FS-JX)in Changsha.FS-JX station and its surrounding 9,117 fire risk points are selected as the fire service supply and demand points,respectively.A total of 196 simulation scenarios throughout a consecutiveweek are analyzed.Eventually,1,933,815 sets of valid sample data are obtained.The results showed that the TECA of FS-JX is 3.27 km^(2),which is far below the standard requirement of 7.00 km^(2) due to the traffic conditions.The visualization results showed that three rivers around FS-JX interrupt the continuity of its effective coverage.The proposed method can provide data support to optimize the locations of fire stations by accurately and dynamically determining the effective coverage of fire stations.

A Framework of LSTM Neural Network Model in Multi-Time Scale Real-Time Prediction of Ship Motions in Head Waves

Ship motions induced by waves have a significant impact on the efficiency and safety of offshore operations.Real-time prediction of ship motions in the next few seconds plays a crucial role in performing sensitive activities.However,the obvious memory effect of ship motion time series brings certain difficulty to rapid and accurate prediction.Therefore,a real-time framework based on the Long-Short Term Memory(LSTM)neural network model is proposed to predict ship motions in regular and irregular head waves.A 15000 TEU container ship model is employed to illustrate the proposed framework.The numerical implementation and the real-time ship motion prediction in irregular head waves corresponding to the different time scales are carried out based on the container ship model.The related experimental data were employed to verify the numerical simulation results.The results show that the proposed method is more robust than the classical extreme short-term prediction method based on potential flow theory in the prediction of nonlinear ship motions.

FPGA implementation of 500-MHz high-count-rate high-time-resolution real-time digital neutron-gamma discrimination for fast liquid detectors

Fast neutron flux measurements with high count rates and high time resolution have important applications in equipment such as tokamaks.In this study,real-time neutron and gamma discrimination was implemented on a self-developed 500-Msps,12-bit digitizer,and the neutron and gamma spectra were calculated directly on an FPGA.A fast neutron flux measurement system with BC-501A and EJ-309 liquid scintillator detectors was developed and a fast neutron measurement experiment was successfully performed on the HL-2 M tokamak at the Southwestern Institute of Physics,China.The experimental results demonstrated that the system obtained the neutron and gamma spectra with a time accuracy of 1 ms.At count rates of up to 1 Mcps,the figure of merit was greater than 1.05 for energies between 50 keV and 2.8 MeV.