Development of a Real-Time RT-PCR for the Universal Detection of LChV1 and Study of the Seasonal Fluctuation of the Viral Titer in Sweet Cherry Cultivars.

摘要: Little cherry virus 1 (LChV1) is a sweet pathogen which has lately been reported in other Prunus spp. LChV1 variability makes reliable detection challenging undertaking. The objective of this work was to develop rapid, sensitive, and one-tube, real-time reverse-transcription polymerase chain reaction (RT-PCR) for the quantification LChV1. Primers TaqMan probe were designed, using conserved regions capsid protein gene. Detection range evaluated several divergent viral isolates. amplification efficiency method estimated at 96.7%, whereas limit about 100 RNA copies. protocol applied study fluctuation within leaves phloem tissue throughout year best periods test plant tissues sample determined. Comparative analysis with previously published nested RT-PCR revealed higher analytical diagnostic sensitivity new test, making it tool that can be used routine testing certification programs.