Comparison of Conventional RT-PCR and Real-time RT-PCR Assays for Diagnosis of Grapevine fanleaf nepovirus (GFLV)*

Abstract

Recently advances in biotechnology and molecular biology have played a significant role in development of serological and molecular assays for detection of plant viruses. However, specificity and sensitivity of them may show variation according to pathogens and hosts. In this study, Grapevine fanleaf nepovirus (GFLV) which was the most prevalent viral agent in Aegean viticulture production areas was used to compare conventional RT-PCR and real-time RT-PCR. For this purpose, 8 infected and 5 healthy, totally 13 symptomatic grapevine samples which have been detected by DAS-ELISA were used. After the conventional RT-PCR and real-time PCR assays, results were evaluated agarose gel electrophoresis and melting analysis respectively. The conventional RT-PCR results showed that 10 out of 13 samples were infected with GFLV, whereas 11 out of 13 samples were found infected with real-time RT-PCR. At the end of this study, the real-time RT-PCR assays were found more sensitive, reliable and rapid than conventional RT-PCR.