Romanian Biotechnological Letters Vol. 22, No. 5, 2017 Copyright © 2017 University of Bucharest Printed in Romania. All rights reserved ORIGINAL PAPER Romanian Biotechnological Letters, Vol. 22, No. 5, 2017 12934 Sensitive Detection of Tomato Spotted Wilt Virus from Pepper Plants by DAS-ELISA, RT-PCR and IC-RT-PCR Received for publication, March 10, 2015 Accepted, September 23, 2016 HANDAN ÇULAL-KILIÇ 1 *, NEJLA YARDIMCI 1 , ALİ BAL 1 , ALİ GÜNEŞ 1 , FATMA DENİZ 1 1 University of SuleymanDemirel University, Agriculture Faculty, Department of Plant Protection, Isparta, Turkey *Address correspondence to: University of SuleymanDemirel University, Agriculture Faculty, Department of Plant Protection,Isparta, Turkey Tel.: +902462114861; Email: handankilic@sdu.edu.tr;handanculal@hotmail.com Abstract 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. Keywords:Pepper, Tomato spotted wilt virus, DAS-ELISA, mechanical inoculation, RT-PCR, IC-RT- PCR. 1. Introduction Pepper (Capsicum annum L.), a member of the Solanaceae family, is an important warm and temperate climate plant.It has been reported that the homeland of pepper is tropical South America, especially Brazil (ANONYMOUS[1]). Pepper was spread from Spain to England in the 1500s and then all over Europe [1].Turkey was the third highest pepper producing country in the world in 2013 with 1.975.269 tons (ANONYMOUS, FAO [2]). In Turkey, 1.975.269 tons of pepper was produced totally (2013) and of this was 879.846 tons of long green pepper, 364.930 tons was sweet bell pepper, 730,493 tons was sauceboat pepper (ANONYMOUS [3]). Isparta, Burdur and Denizli provinces are important for pepper production due to their soil and climatic properties and rich irrigation facilities. In the production of pepper, improper or inadequate agricultural practices and biotic and abiotic disease factors cause significant yield losses. There is a wide variety of fungal, bacterial and viral diseases limiting production. Viral diseases have an important place among these factors due to their chemical and physical structures, size, infection types, symptoms, transportation and a lack of effective management strategies (AGRIOS [4]). In order to minimize the damage caused by viral diseases and to improve the control methods, first it is necessary to identify the viruses found in culture plants. Symptomatologic studies based on the identification of the viruses should be supported by