Establishment of reverse transcription loop-mediated isothermal amplification combined with lateral flow dipstick visual detection method for Actinidia chlorotic ringspot-associated virus

Abstract:

Actinidia chlorotic ringspot-associated virus (AcCRaV) is one of the main viruses that infect kiwifruit. For the rapid detection and monitoring of AcCRaV in kiwifruit, in this study, a reverse transcription loop-mediated isothermal amplification (RT-LAMP) combined with lateral flow dipstick (LFD) method was established, which could achieve rapid and visual detection of AcCRaV in kiwifruit. In this experiment, a set of LAMP specific primers was designed based on the conservative region of the AcCRaV coat protein gene. The optimized reaction system was reacted for 60 min under a constant temperature of 62 ℃, followed by 5 min test strip color reaction to finish the sample detection. The detection sensitivity of the RT-LAMP assay was 100 times higher than that of RT-PCR. The results of RT-LAMP detection of kiwifruit samples were consistent with those by RT-PCR detection, even more, the RT-LAMP results could be visualized through LFD test strips. In conclusion, this method can provide a time-saving and efficient diagnostic tool for detecting AcCRaV infection in kiwifruit.