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

Bacterial wilt caused by is a serious soilborne disease that results in severe losses to tobacco () production in China. In this study, a novel RPA-LFD assay for the rapid visual detection of was established using recombinase polymerase amplification (RPA) and lateral-flow dipstick (LFD). The RPA-LFD assay was performed at 37°C in 30 min without complex equipment. Targeting the sequence of the gene, we designed RPA primers (Rs-rpa-F/R) and an LF probe (Rs-LF-probe) that showed high specificity to . The sensitivity of RPA-LFD assay to was the same as that in conventional PCR at 1 pg genomic DNA, 10 CFU/g artificially inoculated tobacco stems, and 10 CFU/g artificially inoculated soil. The RPA-LFD assay could also detect 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 .