Sensitive Detection of Tomato Spotted Wilt Virus from Pepper Plants by DAS-ELISA, RT-PCR and IC-RT-PCR

This study was conducted to determine the prevalence of Tomato spotted wilt virus (TSWV) in pepper growing areas in Burdur, Isparta and Denizli provinces of Turkey in 2012-2013. During surveys TSWV suspected leaf samples were collected. Total of 448 leaf samples were collected from different locations in the region. Presence of TSWV was investigated using Double antibody sandwich enzyme linked immunosorbent assay (DAS-ELISA) method. ELISA test results showed that 61 samples were infected with TSWV (13.61%). ELISA positive samples for TSWV were used in the mechanical inoculation studies. Typical symptoms of TSWV were observed on the indicator plants. Molecular studies were carried out on 25 ELISA negative samples and 61 ELISA positive samples (Totally 86 samples). 63 samples were detected positive with RT-PCR (reverse transcriptase polymerase chain reaction) method while 70 samples were found to be infected with TSWV using IC-RT-PCR (immunocapture reverse transcriptase polymerase chain reaction) method. Expected size of band with 276 bp was observed on agarose gel electrophoresis. IC-RT-PCR method was found to be more suitable and sensitive than RT-PCR.