Fasulye Tohumlarında Fasulye Adi Mozayik Virüsü’nün Immunocapture RT-PCR ve DAS-ELISA Yöntemleriyle Teşhisi

Year 2022, Volume: 7 Issue: 1, 1 - 7, 29.06.2022

Handan Çulal Kılıç , Ahmet Çat

https://doi.org/10.33484/sinopfbd.1006005

Abstract

Bu çalışmada, 2016 yılında Isparta ve Antalya illerinde fasulye tohumlarında Fasulye adi mozayik virüsü (Bean common mosaic virus: BCMV)’nün belirlenmesi amacıyla yapılmıştır. Bu amaçla araştırma alanından 106 tohum örneği toplanmıştır. Tohum örnekleri biyolojik, serolojik ve moleküler çalışmalar için kullanılmıştır. DAS-ELISA testi sonucunda tohum örneklerinde BCMV’nün hastalık oranı %87.73 olarak belirlenirken, Immunocapture ters transkripsiyon polimeraz zincir reaksiyonu (IC-RT-PCR) yönteminde örneklerdeki hastalık oranı %90.56 olarak belirlenmiştir. Yürütülen çalışmalar sonucunda toplam 96 tohum örneğinde (%90.56) BCMV belirlenmiştir. DAS-ELISA yöntemi ile 93 örnek BCMV ile pozitif belirlenirken; IC-RT-PCR yöntemi ile 96 örnek BCMV ile enfekteli bulunmuştur. IC-RT-PCR yönteminde spesifik primerler ile kılıf protein geninin yaklaşık 850 bp’lik bir kısmı amplifiye edilmiş ve BCMV’ne özgü beklenen seviyede bant elde edilmiştir.

Keywords

BCMV, fasulye, tohum, moleküler teşhis, serolojik method

Detection of Bean Common Mosaic Virus in Bean Seeds by Immunocapture RT-PCR and DAS-ELISA Methods

Abstract

This study was carried out in bean seeds in Isparta and Antalya provinces to determine Bean common mosaic virus (BCMV) in 2016. For this purpose; 106 seed samples were collected from the research area. Seed samples were used for biological, serological, and molecular methods. The virus incidence, as a percentage of seeds samples for BCMV, was found 87.73% by DAS-ELISA and 90.56% by Immunocapture Reverse Transcriptase-Polymerase Chain Reaction (IC-RT-PCR) methods. As a result of the study, BCMV was detected in a total of 96 (90.56%) seed samples. 93 samples were detected positive with the DAS-ELISA method, while 96 samples were found to be infected with BCMV using the IC-RT-PCR method. In the IC-RT-PCR method, approximately 850 bp of the coat protein gene was amplified with specific primers and BCMV specific bands were obtained at the expected levels.

Keywords

BCMV, bean, seed, molecular detection, serological method

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