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Rapid and visual detection of Verticillium dahliae using recombinase polymerase amplification combined with lateral flow dipstick

文献类型： 外文期刊

作者： Ju, Yuliang 1 ; Li, Changfeng 2 ; Shen, Pengfei 1 ; Wan, Ning 1 ; Han, Wenbing 2 ; Pan, Yuemin 1 ;

作者机构： 1.Anhui Agr Univ, Key Lab Biol & Sustainable Management Plant Dis &, Anhui Higher Educ Inst, 130 West Changjiang Rd, Hefei 230036, Anhui, Peoples R China

2.Anhui Acad Agr Sci, Cotton Res Inst, 40 South Nongke Rd, Hefei 230031, Anhui, Peoples R China

关键词： Verticillium dahliae; Recombinase polymerase amplification; Molecular diagnosis; Specificity; Sensitivity; Practicability

期刊名称：CROP PROTECTION （ 影响因子：2.571； 五年影响因子：3.11 ）

ISSN： 0261-2194

年卷期： 2020 年 136 卷

页码：

收录情况： SCI

摘要： Verticillium dahliae is a soil-borne phytopathogenic fungus that causes verticillium wilt disease in cotton, resulting in reduced fiber quality and serious economic losses. However, there is still no efficient management strategies that have been developed to control the disease. Therefore, rapid and accurate detection of V. dahliae would be invaluable in disease control programs. In this study, an assay consisting of recombinase polymerase amplification combined with lateral-flow dipstick technology (RPA-LFD) was developed for the rapid and sensitive detection of V. dahliae. The RPA-LFD assay was completed at the isothermal temperature of 37 degrees C within 25 min. The RPA primers and LF probe exhibited high specificity to V. dahliae. The detection limit of RPA-LFD was 10(3) fg fungal genomic DNA in a 50 mu L reaction volume and 10(3) spores per gram using artificially inoculated soil samples, whereas conventional PCR detected as low as 10(3) fg DNA and 10(4) spores. Additionally, the RPA-LFD assay detected V. dahliae in field soil samples, showing no significant differences compared to conventional PCR. The simple, rapid, and practical nature of the RPA-LFD assay suggests that it will serve as a promising molecular diagnostic tool for the accurate and rapid detection of V. dahliae.

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