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环介导等温扩增技术全面指南,一种用于植物病原真菌的新兴诊断工具。

A comprehensive guide to loop-mediated isothermal amplification, an emerging diagnostic tool for plant pathogenic fungi.

作者信息

Németh Márk Z, Kovács Gábor M

机构信息

Plant Protection Institute, HUN-REN Centre for Agricultural Research, Budapest, Hungary.

Department of Plant Anatomy, Institute of Biology, Eötvös Loránd University, Budapest, Hungary.

出版信息

Front Plant Sci. 2025 May 22;16:1568657. doi: 10.3389/fpls.2025.1568657. eCollection 2025.

DOI:10.3389/fpls.2025.1568657
PMID:40487219
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC12142221/
Abstract

The detection and identification of plant pathogenic fungi are crucial for effective plant protection measures. In the past two decades, loop-mediated isothermal amplification (LAMP) has emerged as a simple and cost-efficient tool for plant disease diagnosis, overcoming many drawbacks of traditional and PCR-based methods. LAMP relies on efficient DNA synthesis at a constant temperature, eliminating the need for thermocycling equipment. It is typically more robust, specific, and sensitive than PCR. This literature review summarizes LAMP primer design, reaction protocol development, sensitivity and specificity testing, and result detection methods. We provide examples of how LAMP's advantages are exploited in disease diagnosis and survey its diverse applications in plant pathogenic fungi research. These applications include the detection, identification, and monitoring of plant pathogenic fungi; the replacement of culture-based methods; the detection of genetic regions associated with functional changes; and the detection of single nucleotide polymorphisms. A comprehensive list of available assays is also provided. Despite its shortcomings-including difficulties with primer design, risks of cross-contamination, and the potential for false positives-LAMP holds significant potential to gain widespread recognition and popularity in the study of plant pathogenic fungi.

摘要

植物病原真菌的检测与鉴定对于有效的植物保护措施至关重要。在过去二十年中,环介导等温扩增技术(LAMP)已成为一种用于植物病害诊断的简单且经济高效的工具,克服了传统方法和基于聚合酶链式反应(PCR)方法的许多缺点。LAMP依靠在恒温下高效的DNA合成,无需热循环设备。它通常比PCR更稳健、特异且灵敏。本文献综述总结了LAMP引物设计、反应体系优化、灵敏度和特异性测试以及结果检测方法。我们举例说明了LAMP的优势在病害诊断中的应用方式,并概述了其在植物病原真菌研究中的多种应用。这些应用包括植物病原真菌的检测、鉴定和监测;基于培养方法的替代;与功能变化相关的基因区域检测;以及单核苷酸多态性检测。还提供了可用检测方法的综合列表。尽管LAMP存在一些缺点,包括引物设计困难、交叉污染风险以及假阳性的可能性,但它在植物病原真菌研究中具有获得广泛认可和普及的巨大潜力。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/289c/12142221/60ba3930ff5a/fpls-16-1568657-g001.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/289c/12142221/60ba3930ff5a/fpls-16-1568657-g001.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/289c/12142221/60ba3930ff5a/fpls-16-1568657-g001.jpg

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A Loop-Mediated Isothermal Amplification Assay for the Rapid Detection of Causing Tea Leaf Spot.
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