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

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