Rapid detection of Banna virus by reverse transcription-loop-mediated isothermal amplification (RT-LAMP)

Abstract

ObjectivesBanna virus (BAV) is classified in the genus Seadornavirus within the Reoviridae family and considered to be an emerging pathogen. We aimed to develop a rapid and simple molecular detection approach for all BAV subgroups in isothermal conditions. MethodA set of six specific primers was designed to target the segment 12 of BAV, and the reverse transcription-loop mediated isothermal amplification (RT-LAMP) assay was developed and compared with conventional RT-PCR method. ResultsThe amplification of the RT-LAMP assay can be obtained within 40min at 65°C. The results from specificity showed that only target BAVs RNA including genotypes A, B and C were amplified and the assay demonstrated a sensitivity of 3.6×10−2PFU/mL, which was higher than conventional RT-PCR measurement. A good reliability for the assay was presented in the further evaluation for BAVs RNA from serial diluted BAV-spiked serum and 47 pools of field mosquito samples. ConclusionsOur findings present a rapid, sensitive and specific RT-LAMP assay that can be applied for BAV detection in clinical or field samples in the future.