Rapid detection of virus based on immunocapture combined with reverse transcription loop-mediated isothermal amplification

Abstract

An immunocapture reverse transcription loop-mediated isothermal amplification (IC-RT-LAMP) was developed for detection of potato virus Y (PVY) and cucumber mosaic virus (CMV). In this procedure, 2 sets of primers were designed to match CMV and PVY specific sequences. The PCR tubes used for this assay were coated with monoclonal antibodies (Mabs) specific for PVY and CMV, respectively. The concentrations of MgSO4 and dNTP of these two detection systems were adjusted and optimized. The assays could be performed in less than 250 minutes, and the results could be identified through trained naked eyes. The use of Mabs combined with RT-LAMP conferred high sensitivity and specificity for virus detection not only in plant tissues, but also in aphids. The detection results between IC-RT-LAMP and RT-PCR were highly consistent. The results showed that IC-RT-LAMP allowed a rapid detection of CMV and PVY in aphids without RNA isolation steps and a thermal cycler. No differences in sensitivity and specificity were detected between IC-RT-LAMP and RT-PCR methods.