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利用重组酶聚合酶扩增-侧向流动试纸条联用技术快速简便地检测草莓中的[具体物质未给出]

Rapid and simple detection of in strawberry using a coupled recombinase polymerase amplification-lateral flow strip assay.

作者信息

Lu Xinyu, Xu Heng, Song Wen, Yang Zitong, Yu Jia, Tian Yuee, Jiang Min, Shen Danyu, Dou Daolong

机构信息

College of Plant Protection, Nanjing Agricultural University, Nanjing, 210095 China.

Department of Plant Protection, Henan University of Science and Technology, Luoyang, 471000 China.

出版信息

Phytopathol Res. 2021;3(1):12. doi: 10.1186/s42483-021-00089-8. Epub 2021 Jun 10.

Abstract

UNLABELLED

is a devastating pathogen that infects a wide range of plants and causes Phytophthora rot disease, which has resulted in great economic losses in crop production. Therefore, the rapid and practicable detection of is important for disease monitoring and forecasting. In this study, we developed a lateral flow recombinase polymerase amplification (LF-RPA) assay for the sensitive visual detection of . Specific primers for were designed based on the ras-related protein gene ; all 10 isolates yielded positive detection results, whereas no cross-reaction occurred in related oomycete or fungal species. The detection limit for the LF-RPA assay was 100 fg of genomic DNA under optimized conditions. Combined with a simplified alkaline lysis method for plant DNA extraction, the LF-RPA assay successfully detected in naturally diseased strawberry samples without specialized equipment within 40 min. Thus, the LF-RPA assay developed in this study is a rapid, simple, and accurate method for the detection of , with the potential for further application in resource-limited laboratories.

SUPPLEMENTARY INFORMATION

The online version contains supplementary material available at 10.1186/s42483-021-00089-8.

摘要

未标记

是一种毁灭性病原体,可感染多种植物并引发疫霉根腐病,给作物生产造成了巨大经济损失。因此,快速且可行的检测对于病害监测和预测至关重要。在本研究中,我们开发了一种侧流重组酶聚合酶扩增(LF-RPA)检测方法,用于对进行灵敏的可视化检测。基于ras相关蛋白基因设计了的特异性引物;所有10个分离株均产生阳性检测结果,而在相关卵菌或真菌物种中未发生交叉反应。在优化条件下,LF-RPA检测方法的检测限为100 fg基因组DNA。结合用于植物DNA提取的简化碱性裂解方法,LF-RPA检测方法在40分钟内无需专门设备即可成功检测自然发病草莓样品中的。因此,本研究开发的LF-RPA检测方法是一种快速、简单且准确的检测方法,具有在资源有限实验室中进一步应用的潜力。

补充信息

在线版本包含可在10.1186/s42483-021-00089-8获取的补充材料。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/972b/8189726/eb605ad532ec/42483_2021_89_Fig1_HTML.jpg

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