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Development of a recombinase polymerase amplification combined with lateral flow dipstick assay for rapid and sensitive detection of bean common mosaic virus

文献类型： 外文期刊

作者： Qin, Jiachao 1 ; Yin, Zhe 3 ; Shen, Danyu 4 ; Chen, Huatao 1 ; Chen, Xin 1 ; Cui, Xiaoyan 1 ; Chen, Xuehao 2 ;

作者机构： 1.Jiangsu Acad Agr Sci, Jiangsu Key Lab Hort Crop Genet Improvement, Inst Ind Crops, Nanjing 210014, Jiangsu, Peoples R China

2.Yangzhou Univ, Coll Hort & Plant Protect, Dept Hort, Yangzhou 225009, Jiangsu, Peoples R China

3.Beijing Plant Protect Stn, Beijing 100029, Peoples R China

4.Nanjing Agr Univ, Coll Plant Protect, Dept Plant Pathol, Nanjing 210014, Jiangsu, Peoples R China

5.Jiangsu Univ, Inst Life Sci, Zhenjiang 212013, Jiangsu, Peoples R China

关键词： Potyvirus; RPA; LFD; Field detection; Viral diagnosis; Phaseolus angularis; Glycine max

期刊名称：PHYTOPATHOLOGY RESEARCH （ 影响因子：3.955； 五年影响因子：3.955 ）

ISSN： 2096-5362

年卷期： 2021 年 3 卷 1 期

页码：

收录情况： SCI

摘要： Bean common mosaic virus (BCMV) is one of the most widespread and damaging viruses of cultivated legumes in the world. In addition to serious yield reduction and germplasm decline, BCMV infection also makes legumes more vulnerable to other pathogens. Early diagnosis of the virus is particularly important in limiting its spread. Recombinase polymerase amplification (RPA) is a novel isothermic amplification technology. The whole reaction can be done outside the laboratory environment after the nucleic acid sample is obtained. In this study, we established a rapid and sensitive RPA combined with the lateral flow dipstick (LFD) assay for detection of BCMV, based on the conserved BCMV coat protein (CP) gene sequence. Specific primers and a probe were designed, which amplify similar to 150bp CP fragments from BCMV-infected samples under a constant temperature of 37 degrees C for 20min. The end-labeled amplification products were detected by high-affinity LFD within 5min. Sensitivity of this RPA-LFD assay was 1000 times greater than that of the conventional polymerase chain reaction (PCR) assay. Furthermore, when the primers/probe were used against related potyviruses including soybean mosaic virus (SMV), bean yellow mosaic virus (BYMV), and turnip mosaic virus (TuMV), the three potyviruses were not detected, indicating that the assay was BCMV species-specific. The RPA-LFD assay was also successfully applied for the detection of seed-borne BCMV in beans. The RPA-LFD assay has great potential application in the rapid diagnosis of BCMV in the field.

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