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鉴定一种秋粘虫(Spodoptera frugiperda)特异性基因,并建立一种快速、敏感的环介导等温扩增检测方法。

Identification of a fall armyworm (Spodoptera frugiperda)-specific gene and development of a rapid and sensitive loop-mediated isothermal amplification assay.

机构信息

Department of Biomedical Gerontology, Hallym University Graduate School, Chuncheon, Gangwon-Do, Republic of Korea.

Ilsong Institute of Life Sciences, Hallym University, Yeongdeungpo-gu, Seoul, Republic of Korea.

出版信息

Sci Rep. 2022 Jan 18;12(1):874. doi: 10.1038/s41598-022-04871-2.

Abstract

The fall armyworm [FAW, Spodoptera frugiperda (J E Smith)], a moth native to America, has spread throughout the world since it was first discovered in Africa in 2016. The FAW is a polyphagous migratory pest that can travel over long distances using seasonal winds or typhoons because of its excellent flying ability, causing serious damage to many crops. For effective FAW control, accurate species identification is essential at the beginning of the invasion. In this study, the FAW-specific gene Sf00067 was discovered by performing bioinformatics to develop a fast and accurate tool for the species-specific diagnosis of this pest. An Sf00067 loop-mediated isothermal amplification (LAMP) assay was developed, and optimal conditions were established. The Sf00067 6 primer LAMP (Sf6p-LAMP) assay established in this study was able to diagnose various genotype-based strains of FAW captured in Korea and FAWs collected from Benin, Africa. Our FAW diagnostic protocol can be completed within 30 min, from the process of extracting genomic DNA from an egg or a 1st instar larva to species determination.

摘要

秋黏虫(FAW,Spodoptera frugiperda (J E Smith))原产于美洲,自 2016 年在非洲首次发现以来已传播到世界各地。FAW 是一种多食性迁徙性害虫,由于其出色的飞行能力,能够利用季节性风或台风远距离迁徙,对许多作物造成严重损害。为了有效防治 FAW,在入侵初期进行准确的物种鉴定至关重要。在这项研究中,通过生物信息学发现了 FAW 特异性基因 Sf00067,开发了一种快速准确的工具,用于该害虫的种特异性诊断。开发了 Sf00067 环介导等温扩增(LAMP)检测方法,并建立了最佳条件。本研究中建立的 Sf00067 6 引物 LAMP(Sf6p-LAMP)检测方法能够诊断在韩国捕获的各种基于基因型的 FAW 菌株和来自非洲贝宁的 FAW。我们的 FAW 诊断方案可以在 30 分钟内完成,从提取卵或 1 龄幼虫的基因组 DNA 到物种鉴定。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/aff6/8766445/a290bcbf5742/41598_2022_4871_Fig1_HTML.jpg

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