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用于检测……的实时比色环介导等温扩增检测方法的开发

Development of Real-Time and Colorimetric Loop Mediated Isothermal Amplification Assay for Detection of .

作者信息

Stehlíková Dagmar, Beran Pavel, Cohen Stephen P, Čurn Vladislav

机构信息

Biotechnological Centre, Faculty of Agriculture, University of South Bohemia, Na Sadkach 1780, 37005 Ceske Budejovice, Czech Republic.

Department of Plant Pathology, The Ohio State University, Columbus, OH 43210, USA.

出版信息

Microorganisms. 2020 Aug 26;8(9):1301. doi: 10.3390/microorganisms8091301.

Abstract

is one of the causal agents of bacterial spot (BS), an economically important bacterial disease of tomato and pepper. Field-deployable and portable loop-mediated isothermal amplification (LAMP)-based instruments provide rapid and sensitive detection of plant pathogens. In order to rapidly and accurately identify and differentiate from other BS-causing spp., we optimized a new real-time monitoring LAMP-based method targeting the -specific gene. Specificity and sensitivity of real-time and colorimetric LAMP assays were tested on the complex of bacterial strains pathogenic to tomato and pepper and on plants infected by the pathogen. The assay detection limit was 1 pg/μL of genomic DNA with an assay duration of only 30 min. The use of portable and handheld instruments allows for fast analysis, reducing the diagnosis time, and can contribute to proper disease management and control of . Due to the high efficiency of this method, we suggest its use as a standard diagnostic tool during phytosanitary controls.

摘要

是番茄和辣椒细菌性斑点病(BS)的致病因子之一,细菌性斑点病是一种对经济有重要影响的番茄和辣椒细菌性病害。基于环介导等温扩增(LAMP)的可现场部署和便携式仪器可对植物病原体进行快速灵敏的检测。为了快速准确地识别并与其他引起细菌性斑点病的物种进行区分,我们针对特定基因优化了一种基于实时监测LAMP的新方法。在对番茄和辣椒致病的复杂细菌菌株以及受病原体感染的植物上测试了实时和比色LAMP检测的特异性和灵敏度。该检测方法的检测限为1 pg/μL基因组DNA,检测时长仅30分钟。使用便携式和手持式仪器可实现快速分析,减少诊断时间,并有助于对[病原体名称]进行适当的病害管理和控制。由于该方法效率高,我们建议在植物检疫控制期间将其用作标准诊断工具。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/29d6/7563391/99fbb89d4220/microorganisms-08-01301-g001.jpg

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