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利用重组酶聚合酶扩增-侧流层析检测快速检测番茄黄曲叶病毒。

A rapid detection of tomato yellow leaf curl virus using recombinase polymerase amplification-lateral flow dipstick assay.

机构信息

Institute of Plant Protection, Key Laboratory of Urban Agriculture (North China), Ministry of Agriculture and Rural Affairs, Beijing Academy of Agriculture and Forestry Sciences, Beijing, China.

State Key Laboratory for Managing Biotic and Chemical Threats to the Quality and Safety of Agro-products, Key Laboratory of Biotechnology in Plant Protection of Ministry of Agriculture and Zhejiang Province, Institute of Plant Virology, Ningbo University, Ningbo, China.

出版信息

Lett Appl Microbiol. 2022 May;74(5):640-646. doi: 10.1111/lam.13611. Epub 2021 Dec 14.

Abstract

Tomato yellow leaf curl disease which is caused by Tomato yellow leaf curl virus (TYLCV) is economically important and a widely spread tomato disease in China. Rapid and accurate detection methods are important in the control TYLCV. Here, a rapid method was developed to identify TYLCV on the basis of recombinase polymerase amplification (RPA) that can be visualized in 5 min using lateral flow dipsticks. The sensitivity and the specificity of this method were evaluated. This method can detect 0·5 pg DNA after 30 min at 37°C without any expensive instrumentation. In addition, it showed higher sensitivity than a PCR method when purified DNA was used. Moreover, the TYLCV was specifically detected, whereas other viruses infecting tomato produced negative results. The crude tomato extracts used in this assay has potential application in minimally equipped plant clinic laboratories. This method will facilitate the early and rapid detection of TYLCV for the timely application of control measures.

摘要

番茄黄曲叶病是由番茄黄曲叶病毒(TYLCV)引起的,在中国是一种经济上重要且广泛传播的番茄病害。快速准确的检测方法对于控制 TYLCV 非常重要。在这里,我们基于重组聚合酶扩增(RPA)建立了一种快速检测方法,使用侧流纸条可在 5 分钟内可视化检测结果。我们评估了该方法的灵敏度和特异性。该方法在 37°C 下孵育 30 分钟后,可检测到 0.5pg 的 DNA,无需任何昂贵的仪器。此外,当使用纯化 DNA 时,该方法的灵敏度高于 PCR 方法。此外,该方法特异性地检测到 TYLCV,而感染番茄的其他病毒则产生阴性结果。本试验中使用的粗提番茄提取物在设备简陋的植物诊所实验室中有潜在的应用价值。该方法将有助于早期和快速检测 TYLCV,以便及时采取控制措施。

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