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不同提取及扩增方法检测大肠杆菌DNA的性能比较

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湘潭大学学报（自然科学版） 2024年第2期46卷 110-122页

作者：余小妹钱纯亘聂立波汤建新吴力强黄钊易辉湖南工业大学生命科学与化学学院生物医用纳米材料与器件湖南省重点实验室湖南株洲412007 深圳市亚辉龙生物科技股份有限公司广东深圳518116 株洲市人民医院湖南株洲412008

大肠杆菌O157∶H7是一种危害性极大的食源性致病菌,为实现大肠杆菌O157∶H7的快速检测,分别采用碱加热裂解法、TritonX-100加热裂解法、溶菌酶加十二烷基硫酸钠(SDS)裂解法、SDS加蛋白酶K裂解法及热裂解法5种方法提取大肠杆菌O157∶H7... 详细信息

大肠杆菌O157∶H7是一种危害性极大的食源性致病菌,为实现大肠杆菌O157∶H7的快速检测,分别采用碱加热裂解法、TritonX-100加热裂解法、溶菌酶加十二烷基硫酸钠(SDS)裂解法、SDS加蛋白酶K裂解法及热裂解法5种方法提取大肠杆菌O157∶H7基因组DNA,考察了环介导等温扩增(LAMP)和实时荧光定量聚合酶链式反应(qPCR)扩增法对所提取DNA的检测效果,并建立最佳的核酸快速提取样品预处理方法.结果表明,以碱加热裂解预处理提取的大肠杆菌O157∶H7基因组DNA为检测样本时,LAMP扩增及qPCR扩增法的检测性能最好,检出限达到10^(1)CFU/mL,与常规商品化试剂盒提取所得菌液基因组DNA的检测性能一致.所建立的碱加热裂解样品预处理方法耗时短、成本低,无须进行复杂的核酸提取,操作简单,可用于大肠杆菌O157∶H7的快速检测.

关键词：大肠杆菌O157∶H7 LAMP qPCR 碱加热裂解法核酸提取

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奶牛乳房炎病原菌研究进展

湖南畜牧兽医 2024年第2期 52-55页

作者：蒋维刘俊琦周望平会同县炮团乡农业综合服务中心湖南会同418399 湖南省畜牧兽医研究所湖南长沙410131

奶牛乳房炎是危害奶牛业健康发展的最重要的疾病之一,该病可引起奶牛产奶量下降,乳汁成分改变,导致奶农收入减少。无乳链球菌、乳房链球菌、停乳链球菌、金黄色葡萄球菌和大肠杆菌是当前奶牛乳房炎最主要的病原菌。文章就奶牛乳房炎在... 详细信息

奶牛乳房炎是危害奶牛业健康发展的最重要的疾病之一,该病可引起奶牛产奶量下降,乳汁成分改变,导致奶农收入减少。无乳链球菌、乳房链球菌、停乳链球菌、金黄色葡萄球菌和大肠杆菌是当前奶牛乳房炎最主要的病原菌。文章就奶牛乳房炎在全球的流行情况、主要致病菌特征进行归纳、概述,以期为日后净化此病,制定科学、合理、有效的综合防制措施提供参考依据。

关键词：奶牛乳房炎链球菌金黄色葡萄球菌大肠杆菌 LAMP

Database system for managing 20,00020-inch PMTs at JUNO

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Nuclear Science and Techniques 2022年第3期33卷 10-21页

作者： Jun Wang Nikolay Anfimov Jing-Yuan Guo Yu Gu Hang Hu Min Li Qiu-Mei Ma Alexander Olshevskiy Zhao-Yuan Peng Zhong-Hua Qin Alexander Tietzsch Bjorn Wonsak Wei Wang Zhi-Min Wang Mei-Hang Xu Wan Xie Zheng-Yun You Hai-Qiong Zhang Rong Zhao School of Physics Sun Yat-Sen UniversityGuangzhou 510275China Joint Institute for Nuclear Research Dubna 141980Russia College of Science and Engineering Ji Nan UniversityGuangzhou 510632China Institute of High Energy Physics Chinese Academy of SciencesBeijing 100049China University of Chinese Academy of Sciences Beijing 100049China State Key Laboratory of Particle Detection and Electronics Beijing 100049China Physikalisches Institut Eberhard Karls Universitat Tubingen72074 TubingenGermany Institut fur Experimentalphysik Universitat Hamburg20148 HamburgGermany Sino-French Institute of Nuclear Engineering and Technology Sun Yat-Sen UniversityZhuhai 519082China

A database system,known as the large PMT characterization and instrumentation database system(LPMT-CIDS),was designed and implemented for the Jiangmen Underground Neutrino Observatory(JUNO).The system is based on a Linux+Apache+MySQL+PHP(LAMP)server and focuses on modularization and architecture separation.It covers all the testing stages for the 20-inch photomultiplier tubes(PMTs)at JUNO and provides its users with data storage,analysis,and visualization services.Based on the successful use of the system in the 20-inch PMT testing program,its design approach and construction elements can be extended to other projects.

关键词： LAMP Photomultiplier tubes Jiangmen underground neutrino observatory Data visualization

Rapid visual detection of Vibrio parahaemolyticus by combining LAMP-CRISPR/Cas12b with heat-labile uracil-DNA glycosylase to eliminate carry-over contamination

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Journal of Zhejiang University-Science B(Biomedicine & Biotechnology) 2023年第8期24卷 749-754页

作者： Fang WU Chen LU Wenhao HU Xin GUO Jiayue CHEN Zhidan LUO Jiangsu Key Laboratory of Marine Biological Resources and Environment Jiangsu Key Laboratory of Marine Pharmaceutical Compound ScreeningJiangsu Ocean UniversityLianyungang 222005China Co-Innovation Center of Jiangsu Marine Bio-industry Technology Jiangsu Ocean UniversityLianyungang 222005China BestEnzymes Biotech Co. Ltd.Lianyungang 222005China

Vibrio parahaemolyticus is a major pathogen frequently found in seafood.Rapid and accurate detection of this pathogen is important for the control of bacterial foodborne diseases and to ensure food safety.In this study,we established a one-pot system that combines uracil-DNA glycosylase(UDG),loopmediated isothermal amplification(LAMP),and clustered regularly interspaced short palindromic repeats(CRISPR)/CRISPR-associated protein 12b(Cas12b)for detecting V.parahaemolyticus in seafood.This detection system can effectively perform identification using a single tube and avoid the risk of carryover contamination.

关键词： LAMP visual system

Development of a loop‑mediated isothermal amplification assay for detection of Austropeplea tomentosa from environmental water samples

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Animal Diseases 2023年第1期3卷 35-48页

作者： Lily Tran Vignesh A.Rathinasamy Travis Beddoe Department of Animal Plant and Soil SciencesSchool of AgricultureBiomedicine and EnvironmentLa Trobe UniversityBundooraVIC 3083Australia Full list of author information is available at the end of the article

Lymnaeid snails are key intermediate hosts for the development and survival of Fasciola spp.,the causative agent of Fascioliasis which are economically important parasites infecting humans and livestock globally.The current control method for treating Fascioliasis is heavily reliant on anthelmintic drugs,particularly Triclabendazole(TCBZ)which has resulted in drug-resistant parasites and poses significant risk as there are no long-term efficacious alternatives available.Sustainable control measures at the farm level could include both parasite and snail control will play an important role in Fasciola spp.control and reduce the reliance on anthelmintic drugs.Implementation of such sustainable control measures requires effective identification of snails on the property however Lymnaeid snails are small and difficult to physically locate.Snail identification using an environmental DNA approach is a recent approach in which physically locating snails are not required.Austropeplea tomentosa,is the primary intermediate snail host for F.hepatica transmission in South-East Australia and we present an in-field loop-mediated isothermal amplification and water filtering method for the detection of A.tomentosa eDNA from water samples to improve current surveillance methods.This methodology is highly sensitive with a detection limit of 5×10^(−6)ng/μL,detected in<20 minutes,with cumulative sample preparation and amplification time under 1 hour.This proposed workflow could assist in monitoring areas to determine the risk of Fascioliasis infection and implement strategies to manage snail populations to ultimately reduce the risk of infection for humans and livestock.

关键词： Fasciola spp. Snail Molecular detection DNA diagnostics LAMP Environmental sampling eDNA

Radiation model of a xenon flash LAMP in a laser amplifier pump cavity

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High Power Laser Science and Engineering 2015年第4期3卷 31-37页

作者： Yongzhong Wu Jianqiang Zhu Zhixiang Zhang Yangshuai Li National Laboratory on High Power Laser and Physics Shanghai Institute of Optics and Fine MechanicsChinese Academy of Sciences University of Chinese Academy of Sciences

Understanding the radiation model of a flash LAMP is essential for the reflector design of a laser amplifier. Reflector design often involves several simplifying assumptions, like a point or Lambertian source; either of these assumptions may lead to significant errors in the output distribution. In practice, source non-idealities usually result in sacrificing the amplifier's gain coefficient. We propose a novel test technique for attaining the xenon flash LAMP absolute spectral intensity at various angles of view, and then accurately predict radiation distributions and generate the reflector shape. It is shown that due to the absorption of emitted radiation by the LAMP itself, the behavior of the radiation model at various wavelengths is different. Numerical results of xenon plasma absorption coefficient were compared with the measured data. A reasonable agreement was obtained for the absorption coefficient parameters. Thus, this work provides a useful analytical tool for the engineering design of laser amplifier reflectors using xenon flash LAMPs as pumps.

关键词： absorption coefficient current density radiation model xenon flash LAMP xenon plasma

Organic and Inorganic Mercury in Biological Samples of Flouresecent LAMP Industries Workers and Health Risks

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Biomedical and Environmental Sciences 2020年第2期33卷 89-102页

作者： Nayab GUL Sardar KHAN Abbas KHAN Javed NAWAB Adnan SARWAR Nida GUL Climate Change Centre The University of AgriculturePeshawar 25120Pakistan Department of Environmental Sciences University of PeshawarPeshawar 25120Pakistan Drug Control Administration Government of Khyber PakhtunkhwaHealth DepartmentPeshawar 25120Pakistan Department of Environmental Sciences Abdul Wali Khan University MardanPeshawar 25120Pakistan Pak International Medical College and Research Institute HayatabadPeshawar 25120Pakistan

Objective The present study aims to investigate the concentrations of Hg and its aspects methyl mercury(Me-Hg)and inorganic mercury(I-Hg)in the biological samples(BSs)of fluorescent LAMP industries workers(FLIWs).Methodology Different BSs including red blood cells(RBCs),plasma,urine,hair and nails were collected from the workers exposed to Hg and unexposed persons were selected as control group to measure both the T-Hg concentration as well as its species in different biological samples through quantitative analysis.Health data was collected through questionnaire survey.Results The mean concentrations of T-Hg(31.9μg/L),Me-Hg(27.7μg/L),and I-Hg(5.36μg/L)in RBCs were found significantly(P<0.001)higher among the workers(n=40)as compared to the control group(n=40).Similarly the mean Hg concentrations in plasma,urine,hair and nails were also significantly higher among the workers than the control group.The statistical relation between Hg concentration and demographic characteristics observed that workers experience and fish consumption has increased the Hg concentration while age,weight and smoking found no significant effect on Hg concentration in the BSs.Conclusion The study observed that the workers were highly exposed to high concentration of Hg and they are at a high health risk.

关键词： Mercury Health risk Fluorescent LAMP industries Regression Demographic characteristics Biological samples

Nucleic acid-based diagnostics for infectious diseases in public health affairs

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Frontiers of Medicine 2012年第2期6卷 173-186页

作者： Albert Cheung-Hoi Yu Greg Vatcher Xin Yue Yan Dong Mao Hua Li Patrick H.K.Tam Parker Y.L.Tsang April K.Y.Wong Michael H.K.Hui Bin Yang Hao Tang Lok-Ting Lau Neuroscience Research Institute/Department of Neurobiology School of Basic Medical Sciences/Key Laboratory for NeuroscienceMinistry of Education/Key Laboratory for NeuroscienceMinistry of HealthPeking UniversityBeijing 100191China Hai Kang Life Corporation Limited Hong Kong SARChina and Beijing 100176China

Infectious diseases,mostly caused by bacteria and viruses but also a result of fungal and parasitic infection,have been one of the most important public health concerns throughout human history.The first step in combating these pathogens is to get a timely and accurate diagnosis at an affordable cost.Many kinds of diagnostics have been developed,such as pathogen culture,biochemical tests and serological tests,to help detect and fight against the causative agents of diseases.However,these diagnostic tests are generally unsatisfactory because they are not particularly sensitive and specific and are unable to deliver speedy results.Nucleic acid-based diagnostics,detecting pathogens through the identification of their genomic sequences,have shown promise to overcome the above limitations and become more widely adopted in clinical tests.Here we review some of the most popular nucleic acid-based diagnostics and focus on their adaptability and applicability to routine clinical usage.We also compare and contrast the characteristics of different types of nucleic acid-based diagnostics.

关键词： nucleic acid-based diagnostics infectious disease PCR NASBA LAMP microarray LOAC public health affairs

Using knowledge inference to suppress the LAMP disturbance for fire detection

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Journal of Safety Science and Resilience 2021年第3期2卷 124-130页

作者： Kailai Sun Qianchuan Zhao Xinwei Wang Center for Intelligent and Networked Systems Department of AutomationBNRistTsinghua UniversityBeijingChina

Fire detection in buildings is crucial for people’s lives and property.Conventional temperature and smoke sen-sors have many disadvantages:the limited cover range;detection delays;the difficulty in distinguishing smoke and fire.Recently,research on convolutional neural networks(CNN)for fire image detection has become a hot topic.However,existing fire classification and object detection methods are often interfered with by flash-lights,red objects and the high-brightness background,resulting in a high false alarm rate.Besides,light and LAMPs often exist in buildings.To address this issue,this paper focuses on introducing scene prior knowledge and causal inference mechanisms to suppress the LAMP disturbance.Firstly,we train the YoloV3 network to detect and recognize LAMPs.Secondly,to reduce the dataset bias,we mask the LAMP regions with the pro-posed Local Grabcut segmentation method.Last,compared with direct fire classification methods,our proposed methods reduce about 34.6%false alarm rate based on InceptionV4 networks.The experimental results verify the effectiveness among different CNN architectures(Resnet101,Firenet,Densenet121).The code is online at https://***/kailaisun/fire-detection-without-LAMP.

关键词： fire detection LAMP prior knowledge causal inference convolutional neural networks