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Chinese Chemical Letters 2024年第9期35卷 262-265页

作者： Yanqi Wu Yuhong Guan Peilin Huang Hui Chen Liping Bai Zhihong Jiang State Key Laboratory of Quality Research in Chinese Medicine Macao University of Science and TechnologyMacao 999078China Shenzhen LemnisCare Medical Technology Co. Ltd.Shenzhen 518107China Hunan Key Laboratory of Biomedical Nanomaterials and Devices Hunan University of TechnologyZhuzhou 412077China

Norovirus is an infectious disease that can cause non-bacterial gastroenteritis,which has a low infectious dose,rapid onset,and strong transmission ability;therefore,rapid and sensitive detection is essential to reduce the transmission of *** the study,a norovirus GII loop-mediated isothermal amplification assay was developed and prepared into freeze-drying microspheres,and a closed-cassette-based,integrated,reagent-ambient storage,on-site instant detection platform for norovirus GII was constructed using a commercial,fully automated nucleic acid analyzer with integrated magnetic bearing based nuclear acid extraction and nucleic acid detection,with a sensitivity of 10 copies/μL,with no cross-reactivity with other 5 *** 28 simulated samples,the integrated assay platform was consistent with the experimental results of reverse transcription-quantitative polymerase chain reaction(RT-qPCR)assays after conventional laboratory nucleic acid *** entire process can be finished in about 1 h,which is ideal for immediate rapid detection.

关键词： Freeze-dried microspheres Point-of-care testing loop-mediated isothermal amplification Norovirus GII Integrated testing

Rapid detection of the E198A mutation of carbendazim-resistant isolates in Colletotrichum gloeosporioides by loop-mediated isothermal amplification

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Horticultural Plant Journal 2022年第3期8卷 289-296页

作者： Hongbo Yuan Hui Hou Zengqiang Zhou Hongtao Tu Li Wang Zhengzhou Fruit Research Institute Chinese Academy of Agricultural SciencesZhengzhou 450009China

Apple bitter rot is a serious agricultural disease caused by Colletotrichum *** recent years,carbendazim-resistant *** strains bearing an E198A point mutation in theβ-tubulin gene(GAG to GCG)have emerged,threatening global apple *** such,rapidly detecting the presence of this E198Amutation in *** isolates is essential in order to monitor the spread of this pathogen and to prevent outbreaks of ***,we developed a simple loop-mediated isothermal amplification(LAMP)approach to detecting the E198A mutation in *** isolates from‘Gala’apple *** optimized LAMP protocol was sufficient to establish the E198A genotype of a given isolate following a 60min incubation at 63℃ by using four specific *** results of this reaction could be interpreted visually based on a fluorescent yellow-green color change upon the addition of the SYBR Green I dye,and were additionally confirmed via gel ***,this LAMP assay was capable of rapidly and reliably detecting apples that were infected with carbendazim-resistant isolates harboring this E198A *** conclusion,this LAMP assay in this study can rapidly,specifically,and sensitively detect cases of apple bitter rot caused by *** isolates harboring the E198A mutation.

关键词： Apple bitter rot Colletotrichum gloeosporioides Carbendazim resistance loop-mediated isothermal amplification E198A mutation

Quantum Dot Nanobeads-Labelled Lateral Flow Immunoassay Strip for Rapid and Sensitive Detection of Salmonella Typhimurium Based on Strand Displacement loop-mediated isothermal amplification

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Engineering 2022年第12期19卷 62-70页

作者： Yuting Shang Shuzhen Cai Qinghua Ye Qingping Wu Yanna Shao Xiaoying Qu Xinran Xiang Baoqing Zhou Yu Ding Moutong Chen Liang Xue Honghui Zhu Jumei Zhang Guangdong Provincial Key Laboratory of Microbial Safety and Health&State Key Laboratory of Applied Microbiology Southern China&Institute of Microbiology Guangdong Academy of SciencesGuangzhou 510070China State Key Laboratory of Food Science and Technology&National Engineering Research Center for Functional Foods&Synergetic Innovation Center of Food Safety&Joint International Research Laboratory on Food Safety School of Food Science and TechnologyJiangnan UniversityWuxi 214122China

Rapid,sensitive,point-of-care detection of pathogenic bacteria is important for food *** this study,we developed a novel quantum dot nanobeads-labelled lateral flow immunoassay strip(QBs-labelled LFIAS)combined with strand displacement loop-mediated isothermal amplification(SD-LAMP)for quantitative Salmonella Typhimurium(ST)*** dot nanobeads(QBs)served as fluorescence reporters,providing good detection *** customizable strand displacement(SD)probe was used in LAMP to improve the specificity of the method and prevent by-product *** was based on a sandwich immunoassay.A fluorescence strip reader measured the fluorescence intensity(FI)of the test(T)line and control(C)*** linear detection range of the strip was 10^(2)–10^(8) colony forming units(CFU)·mL^(-1).The visual limit of detection was 10^(3) CFU·mL^(-1),indicating that the system was ten-fold more sensitive than AuNPs-labelled test *** specificity was analyzed in accordance with agarose gel outputs of polymerase chain reaction(PCR)and *** detected ST in foods with an acceptable recovery of 85%–110%.The method is rapid,simple,almost equipment-free,and suitable for bacterial detection in foods and for clinical diagnosis.

关键词： Salmonella Typhimurium Quantum dot nanobeads Lateral flow immunoassay strip loop-mediated isothermal amplification Strand displacement probe

Gold Nanoparticles Based Colorimetric Detection of Target DNA After loop-mediated isothermal amplification

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Chemical Research in Chinese Universities 2013年第3期29卷 424-428页

作者： ZHOU Chao MU Ying YANG Meng-chao WU Qing-qing XU Wei ZHANG Ying JIN Wei SONG Qi WU Zhong-yu JIN Qin-han Research Center for Analytical Instrumentation Institute of Cyber-Systems and Control State Key Laboratory of Industrial Control Technology Zhejiang University Hangzhou 310058 P. R. China Central Laboratory Affiliated Hospital of Guiyang Medieal College Guiyang 550004 P. R. China School of Mathematics and Science Shijiazhuang University of Economics Shijiazhuang 050031 P. R. China

We have developed a rapid, simple and label-free colorimetric method for the identification of target DNA It is based on loop-mediated isothermal amplification（LAMP）. Plain gold nanoparticles（AuNPs） are used to indicate the occurrence of LAME The amplified product is mixed with AuNPs in an optimized ratio, at which the deoxyribonucleotides（dNTPs） bind to the AuNPs via ligand-metal interactions and thus enhance AuNPs stability. If a target DNA is amplified, the dramatic reduction of the dNTPs leads to the aggregation of AuNPs and a color change from red to blue. The success of the method strongly depends on the ionic strength of the solution and the initial concentration of dNTPs. Unlike other methods for the identification of isothermal products, this method is simple and can be readily applied on site where instrumentation is inadequate or even lacking.

关键词： Label-free Gold nanoparticle loop-mediated isothermal amplification Naked eyes

Rapid,specific and sensitive detection of Vibrio vulnificus by loop-mediated isothermal amplification targeted to vvhA gene

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Acta Oceanologica Sinica 2018年第4期37卷 83-88页

作者： ZHANG Lina WANG Mingyi CONG Dianxia DING Shuyan CONG Rinan YUE Jinyong GENG Jianli HU Chengjin Department of Laboratory Medicine General Hospital of Jinan Military Region of PLA Department of Central Laboratory Weihai Municipal Hospital affiliated to Dalian Medical University

Vibrio vulnificus is an estuarine bacterial pathogen for *** rapid,specific and sensitive detection of *** is urgently needed for early disease diagnosis and timely treatment of *** *** the study,a loop-mediated isothermal amplification（LAMP） technique was developed for *** detection with a set of primers,composed of two out primers and two inner primers targeted to vvh A *** optimal amplification temperature was 63°C and the reaction only took 35 *** amplification products could not only be detected by agarose gel electrophoresis with ladder-like pattern bands,but also could be visualized using calcein with naked eye ***-five strains were tested for the specificity of LAMP assay,and all the *** strains were identified correctly while other strains were negative *** sensitive of the new LAMP assay was 100-fold more sensitive than the conventional ***,all the *** strains were detected correctly in spiked,clinical and environmental samples by the new LAMP *** with other well-known techniques,the new LAMP assay targeted to vvh A gene was extremely rapid,simple,sensitive and specific for *** identification.

关键词： Vibrio uulnificus loop-mediated isothermal amplification vvhA gene

Direct Detection of Target Genes in Viable Bacteria and Extracellular DNA Using loop-mediated isothermal amplification Assay

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渔业科学进展 2020年第2期41卷 41-50页

作者： YANG Qianqian ZHANG Xuzhi JIANG Xiaoyu LI Yang ZHAO Jun HAO Zhihui WANG Pingping QU Keming College of Marine Sciences Shanghai Ocean University Yellow Sea Fisheries Research Institute Chinese Academy of Fishery Sciences School of Chemistry and Pharmaceutical Sciences Qingdao Agriculture University

When the loop-mediated isothermal amplification(LAMP) assay is used for detecting target genes, DNA extraction is unnecessary in many cases. Simple pretreatment(e.g. heating) is enough to obtain rather sensitive responses. Even test samples without any pretreatment can be used as template. This feature suggests that LAMP is superior to PCR in developing point-of-care test strategies. In this study, using Stx1 gene from E. coli as model, we verified that viable cells, dead cells and extracellular DNA could function as template in the LAMP assay. In the incubation at 63℃, viable bacteria in the LAMP reaction mixture lysed completely within 2 min, providing DNA template for nucleic acid amplification. The Stx1 gene in diluted culture medium, spiked tap water, spiked seawater and real seawater all could be detected, with or without the step of DNA extraction. We found that the complex substances in real sample(e.g. natural seawater) exhibited considerable inhibitory effect on the sensitivity of the LAMP assay. These outcomes are meaningful for building a point-of-care strategy by employing the LAMP assay for environmental monitoring, bio-resource surveys, food safety, etc. in particular those based on environmental DNA.

关键词： loop-mediated isothermal amplification DNA extraction-free Direct gene detection Viable cell Extracellular DNA

Preliminary investigation and detection based on loop-mediated isothermal amplification(LAMP)of phytoplasmas associated with diseases in *** L.

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Oil Crop Science 2022年第4期7卷 219-224页

作者： Yancheng Wen Shufen Zhang Junping He Dongfang Cai Jiacheng Zhu Jianping Wang Jinhua Cao Kun Hu Lei Zhao Dongguo Wang Yizi Liu Institute of Industrial Crops of Henan Academy of Agricultural Sciences Zhengzhou450002China

In the last decade,some disease occurred on our experimental farms that had caused serious *** were not caused by fungi,bacteria or *** loop-mediated isothermal amplification(LAMP)technique,the detection results pointed to the possible pathogen as *** investigation results implied that phytoplasmas could cause more than 13 kinds of symptoms in almost all parts of plants in *** L.,including witches’broom,multi-stems,aggregate main inflorescences,and flat *** incidences of these phytoplasma-associated diseases in our experimental farms rose from 1.61%in 2010 to 6.00%in *** phytoplasma infected plants died without any growing *** studies would be helpful for detecting phytoplasmas diseases,selecting disease resistant germplasm and improving varieties with disease resistances in *** L.

关键词： B.napus L. Phytoplasma associated disease Pathogen Detection loop-mediated isothermal amplification (LAMP)

Rapid and Visualized Detection Method of Canine Parvovirus Using loop-mediated isothermal amplification

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Animal Husbandry and Feed Science 2010年第1期2卷 34-37页

作者： YANG Hui QU Guang-gang ZHAO Yuan-kai SHEN Zhi-qiang Shandong LuDu Bio-science Industry Co. Ltd. Binzhou 256600 China Shandong Binzhou Animal Science and Veterinary Medicine Academy Binzhou 256600 China

[Objective] To develop a rapid and visualized detection method using loop-mediated isothermal amplification （LAMP）, improve the detection rate of canine parvovirus （CPV） and reduce the testing cost. [ Method] According to the conserved regions of the VP2 gene of CPV, six primers were designed to amplify the special DNA sequences by LAMP. In addition, the reaction conditions of LAMP were optimized, and the sensitivity, specificity, repeatability and stability were verified. [ Result] The optimal reaction time of the LAMP method for CPVwas 60 min. The products obtained by LAMP had high specificity without cross-reaction with other generic viruses. The sensitivity of the LAMP was 100 times higher than that of PCR. [ Conclusion] The LAMP method for detecting CPV has high practical value. It has many advantages such as high specificity, high sensitivity, simple operation, low cost and rapid analysis, and it does not require special equipment. Therefore, this method is more suitable for the detection of CPV.

关键词： Canine parvovirus loop-mediated isothermal amplification Diagnosis method

Detection of helminths by loop-mediated isothermal amplification assay:a review of updated technology and future outlook

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Infectious Diseases of Poverty 2019年第2期8卷 92-92页

作者： Miao-Han Deng Lan-Yi Zhong Okanurak Kamolnetr Yanin Limpanont Zhi-Yue Lv Zhongshan School of Medicine Sun Yat-sen UniversityGuangzhou 510080China Faculty of Tropical Medicine Mahidol UniversityBangkok 10400Thailand 不详

Background:Helminths are endemic in more than half of the world's countries,raising serious public health *** diagnosis of helminth infection is crucial to control *** parasitological methods,serological tests and PCR-based assays are the major means of the diagnosis of helminth infeaion,but they are time-consuming and/or expensive,and sometimes provide inaccurate *** mediated isothermal amplification(LAMP)assay,a sensitive,simple and rapid method was therefore developed for deteaion of *** study aims to discuss the current status of application of LAMP on helminths detection and to make a comprehensive evaluation about this updated technology and its future outlook by comparing with several other diagnostic *** body:This review summarizes LAMP assay applied for helminth detection and helminthiasis *** basic principle of LAMP is introduced to help better understand its characteristics and each reported assay is assessed mainly based on its detection sensitivity,specificity and limitations,in comparison with other common diagnostic ***,we discuss the limitations of the assays so as to clarify some potential ways of ***:Here,we summarize and discuss the advantages,disadvantages and promising future of LAMP in heliminth detection,which is expected to help update current knowledge and future perspectives of LAMP in highly sensitive and specific diagnosis and surveillance of helminthiasis and other parasitic diseases,and can contribute to the elimination of the diseases from endemic areas.

关键词： loop-mediated isothermal amplification Helminth Point-of-care-test Epidemiological surveillance Field survey

Establishment of a loop-mediated isothermal amplification Method for Rice Bacterial Leaf Brown Spot Disease

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Journal of Northeast Agricultural University(English Edition) 2023年第1期30卷 13-19页

作者： Zhang Jun-hua Wang Liang Zhang Yao Ni Zhe Xu Xiao-feng Yang Ming-xiu Peng Li-li Yang Xin Wang Yi-han Jiang Xiao-jiao Haseeb Younis College of Agriculture Northeast Agricultural UniversityHarbin 150030China

Rice bacterial leaf brown spot disease caused by Pseudomonas syringae ***(Pss)is a major disease on *** recent years,Pss has emerged worldwide,seriously affecting rice *** is very important to establish a rapid detection method of Pss for the diagnosis and prevention of this *** order to robust and accurately diagnose the rice bacterial leaf brown spot disease in the field and laboratory,an assay system for the Pss was developed in this study,and the specific sequence of hrcN was used as the target,based on loop-mediated isothermal amplification(LAMP).The best detection system was MgSO 48 mmol·L^(-1),Bst DNA polymerase 8 U,dNTP 1.4 mmol·L^(-1),the ratio of internal and outer primers was 2:1,the reaction temperature was 63℃,the reaction time was 45 min,and the lowest sensitivity was 104 CFU·mL^(-1).This results provided an accurate and robust method for laboratory and field diagnosis of bacterial leaf brown spot disease of rice.

关键词： Pseudomonas syringae pv.syringae loop-mediated isothermal amplification a rapid detection method