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Development of Recombinase Polymerase Amplification Combined with Lateral Flow Detection Assay for Rapid and Visual Detection of Ralstonia solanacearum in Tobacco

文献类型： 外文期刊

作者： Li, Changfeng 1 ; Ju, Yuliang 2 ; Shen, Pengfei 2 ; Wu, Xun 2 ; Cao, Le 2 ; Zhou, Benguo 3 ; Yan, Xiaoming 1 ; Pan, Yuemin 2 ;

作者机构： 1.Anhui Acad Agr Sci, Cotton Res Inst, Hefei 230031, Anhui, Peoples R China

2.Anhui Agr Univ, Key Lab Biol & Sustainable Management Plant Dis &, Anhui Higher Educ Inst, Hefei 230036, Peoples R China

3.Anhui Acad Agr Sci, Inst Tobacco, Hefei 230031, Anhui, Peoples R China

关键词： Ralstonia solanacearum; recombinase polymerase amplification; molecular diagnosis; specificity; sensitivity

期刊名称：PLANT DISEASE （ 影响因子：4.614； 五年影响因子：5.33 ）

ISSN： 0191-2917

年卷期： 2021 年 105 卷 12 期

页码：

收录情况： SCI

摘要： Bacterial wilt caused by Ralstonia solanacearum is a serious soilborne disease that results in severe losses to tobacco (Nicotiana tabacum) production in China. In this study, a novel RPA-LFD assay for the rapid visual detection of R. solanacearum was established using recombinase polymerase amplification (RPA) and lateral-flow dipstick (LFD). The RPA-LFD assay was performed at 37 degrees C in 30 min without complex equipment. Targeting the sequence of the RipTALI-9 gene, we designed RPA primers (Rs-rpa-F/R) and an LF probe (Rs-LF-probe) that showed high specificity to R. solanacearum. The sensitivity of RPA-LFD assay to R. solanacearum was the same as that in conventional PCR at 1 pg genomic DNA, 10(3) CFU/g artificially inoculated tobacco stems, and 10(4) CFU/g artificially inoculated soil. The RPA-LFD assay could also detect R. solanacearum from plant and soil samples collected from naturally infested tobacco fields. These results suggest that the RPA-LFD assay developed in this study is a rapid, accurate molecular diagnostic tool with high sensitivity for the detection of R. solanacearum.

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