A reverse transcription-cross-priming amplification method with lateral flow dipstick assay for the rapid detection of Bean pod mottle virus

Qian-Qian Yang,Dao Wang,Xing-Xing Zhao,Xiao-Ping Yu,Peng-Jun Zhang,Jing-Yuan Liu,Shuang Wei,Zi-Hong Ye,Xue-Nan Hu

doi:10.1038/s41598-021-03562-8

Qian-Qian Yang, Dao Wang + Show 7 more

Open Access

https://doi.org/10.1038/s41598-021-03562-8

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Abstract

Bean pod mottle virus (BPMV) is a destructive virus that causes serious economic losses in many countries every year, highlighting the importance of its effective detection. In this study, we developed a fast reverse transcription-cross-priming amplification (RT-CPA) coupled with lateral flow dipstick (LFD) diagnostic method for BPMV detection. The RT-CPA-LFD assay that targets the coat protein gene of BPMV was highly specific against diagnosing four other common viruses transmitted by soybean seeds, i.e., Southern bean mosaic virus (SBMV), Tomato ringspot virus (ToRSV), Arabis mosaic virus (ArMV), and Tobacco ringspot virus (TRSV). The sensitivities of the real-time fluorescent RT-CPA and the RT-CPA-LFD assay were at least 50 pg/μl and 500 pg/μl, respectively. Despite a compromise in the limit of detection of the RT-CPA method compared with TaqMan-MGB real-time RT-PCR, our results demonstrated a notably better performance in the detection of field samples of BPMV-infested soybean seeds. With the advantages of efficiency and convenience by visual determination, the RT-CPA-LFD assay presents a potential application for the rapid and accurate detection of BPMV in routine tests.

Highlights

Bean pod mottle virus (BPMV) is a destructive virus that causes serious economic losses in many countries every year, highlighting the importance of its effective detection
Soybean seeds imported from the USA, which had suspected BPMV infection with seed coat mottling or bleeding hilum symptoms that were captured by Guangzhou Customs District of China, were used as field samples
Primer screening based on real‐time fluorescent reverse transcription (RT)‐CPA for BPMV detection

Summary

Introduction

Bean pod mottle virus (BPMV) is a destructive virus that causes serious economic losses in many countries every year, highlighting the importance of its effective detection. Despite a compromise in the limit of detection of the RT-CPA method compared with TaqMan-MGB real-time RT-PCR, our results demonstrated a notably better performance in the detection of field samples of BPMV-infested soybean seeds. Bean pod mottle virus (BPMV) is a comovirus that has a bipartite positive-strand RNA genome consisting of RNA-1 and RNA-21,2. It was first described on the common bean Phaseolus vulgaris L. var. China, which is one of the largest importers of soybeans, has listed BPMV among the plant quarantine pests due to its serous damage.

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