Reverse transcription loop-mediated isothermal amplification (RT-LAMP) assay for rapid diagnosis of Sugarcane yellow leaf virus in sugarcane

Abstract

Loop mediated isothermal amplification (LAMP) is a rapid, highly specific and reliable technique, which is widely incorporated in the field for diagnosis of pathogens at their early stage of infection. In present study, RT-LAMP (Reverse transcription- Loop mediated isothermal amplification) assay was developed successfully to detect Sugarcane yellow leaf virus (SCYLV) causing yellow leaf disease (YLD) in sugarcane and compared with conventional reverse transcription polymerase chain reaction (RT-PCR) and quantitative real time (qRT-PCR) to assess its sensitivity. RT-LAMP was able to detect the SCYLV up to a limit of 10 pg (picogram), while, RT-PCR assays were able to detect this virus up to 100 pg only. Similarly, RT-LAMP was able to detect up to a limit of 10 pg in the sugarcane cultivar (Co 06030) showing low virus titre whereas, in the same cultivar, RT-PCR and qRT-PCR showed a detection limit of 100 pg. This study revealed high sensitivity of RT-LAMP in detecting SCYLV in sugarcane and this is the first study to be reported on RT-LAMP based diagnosis of SCYLV in Indian sugarcane cultivars.