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

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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