Development and application of Recombinase Polymerase Amplification assay for detection of Bipolaris sorokiniana
Wei Zhao, Yuankai Chi, Meng-di Ye, Tao Wang, Amei Xu, Rende Qi
Abstract
Bipolaris sorokiniana is a fungal pathogen that caused root rot and spot blotch on wheat. A Recombinase Polymerase Amplification (RPA) assay was developed to detect B. sorokiniana specifically based on the sequences of calmodulin gene sequence. The specificity of RPA assay was tested against 19 fungal organisms associated with wheat, and the detection limit was determined to be 10 pg for B. sorokiniana pure fungal DNA. Further, the RPA assay was capable of detecting B. sorokiniana from artificially inoculated and field collected wheat tissues directly. These results indicate that the RPA assay is rapid and effective method for detection and molecular identification of B. sorokiniana.