Loop-mediated isothermal amplification for rapid detection of the causal agents of cassava brown streak disease

The causal agents of cassava brown streak disease have recently been identified as Cassava brown streak virus (CBSV) and Ugandan cassava brown streak virus (UCBSV). Primers have been developed for rapid detection of these viruses by reverse transcription loop-mediated isothermal amplification (RT-LAMP). Performance of the RT-LAMP assays compared favourably with published RT-PCR and real-time RT-PCR methods. Furthermore, amplification by RT-LAMP is completed in 40 min and does not require thermal cycling equipment. Modification of the RT-LAMP reactions to use labelled primers allowed rapid detection of amplification products using lateral flow devices containing antibodies specific to the incorporated labels, avoiding the need for fluorescence detection or gel electrophoresis.

► RT-LAMP primers were developed for detection of CBSV and UCBSV in cassava. ► Sensitivity of the RT-LAMP assays approached that of real-time RT-PCR. ► Amplification was observed by real-time fluorescence monitoring in under 40 min. ► Labelled RT-LAMP products were also detected using lateral flow devices.