RFLP typing of infectious bursal disease virus strains detected in Turkey

Abstract

In this study, identification of IBDVs encountered in Turkey by agar gel immunodiffusion (AGID) and reverse transcription/polymerase chain reaction (RT/PCR) test and molecular typing of IBDV strains by restriction fragment length polymorphism (RFLP) assay were aimed. For this reason, a total of 122 bursa Fabricii from 24 flocks showing the symptoms of IBD, reared in the restricts of Adana, Bolu, Bursa, Çorum and Eskişehir were investigated for the existance of viral antigens and the viral RNAs by AGID and RT/PCR tests, respectively. Molecular typing of RT/PCR positive IBDV samples by RFLP analysis using BstNI, MboI and SspI restriction enzymes was performed. After AGID tests, 102 (83.6 %) of 122 bursal samples from 24 flocks were found to have IBDV antigens. After RT/PCR tests, 47 (38.5 %) out of 122 bursa samples were found to have IBDV RNA while 75 (61.5 %) were not. Following RFLP analysis of RT/PCR positive samples, two different molecular patterns were detected. Following the restriction endonuclease analysis performed with SspI enzyme, these were found to have similar molecular patterns with the European very virulent IBDVs (vvIBDVs). As a conclusion, AGID test was found to be an appropriate test for the routine identification of IBDV strains from bursa samples and RT/PCR-RFLP test could be used for the molecular typing of IBDV virus strains in Turkey

Keywords

• AGID
• IBDV
• molecular typing
• RT/PCR-RFLP

Türkiye’de saptanan infeksiyöz bursal hastalık virus suşlarının RFLP ile tiplendirilmesi

Abstract

Bu çalışmada, Türkiye’de İnfeksiyöz Bursal Hastalığa (IBD) neden olan virusların agar jel immunodifuzyon testi (AGID) ve reverz transkripsiyon/polimeraz zincir reaksiyonu (RT/PCR) ile saptanması ve IBDV suşlarının RFLP (restriction fragment length polymorphism) analizi ile moleküler tiplendirilmesi amaçlandı. Bu amaçla, çeşitli bölgelerdeki 24 sürüye ait toplam 122 adet bursa Fabricius materyali IBDV antijenlerinin varlığı yönünden AGID testi, IBDV viral RNA’sının varlığı yönünden ise RT/PCR testi ile değerlendirildi. RT/PCR sonucunda pozitif olduğu belirlenen virus örneklerinin moleküler tiplendirilmesi, BstNI, MboI ve SspI enzimlerinin kullanıldığı RFLP tekniği ile yapıldı. AGID testleri sonucunda incelenen 24 sürüye ait toplam 122 adet örneğin 102 (% 83.6)’si IBDV yönünden pozitif bulundu. Bursa Fabricius örneklerinden izole edilen RNA’lara uygulanan RT/PCR testi sonucunda, 24 sürüye ait toplam 122 adet materyalin 47 (% 38.5)’sinde viral genetik materyal saptandı. RT/PCR testiyle saptanan IBDV suşlarının RFLP analizi sonucunda birbirinden farklı moleküler yapı gösteren iki farklı virus suşu belirlendi. Bu suşların SspI enzimi ile incelenmesi sonucunda bunların Avrupa’da gözlenen yüksek virulensteki suşlarla benzer moleküler yapıya sahip oldukları ortaya kondu. Sonuç olarak, AGID testinin IBDV suşlarının rutin identifikasyonu için uygun bir test olduğu, RT/PCRRFLP testinin ise IBDV viruslarının tiplendirilmesinde kullanılabileceği belirlendi

Keywords

• AGID
• IBDV
• moleküler tiplendirme
• RT/PCR-RFLP

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