RT-PCR amplification and cloning of both genome segments of an Indian infectious bursal disease virus

Infectious bursal disease virus (IBDV) causes severe immuno-suppression in the young chickens leading to concurrent infections with opportunistic pathogen. To characterize Indian field isolate of IBDV at molecular level, the complete coding sequence of genome segments A and B of IBDV has been amplified in to three (A1–1.2 kb, A2–1.3 kb and A3–1.3 kb) and four (B1–718 bp, B2–712 bp, B3–717 bp and B4–740 bp) overlapping fragments, respectively by RT-PCR. The authenticity of the amplicons was verified by their size in agarose gel. The fragments thus generated could be cloned in to pGEMT vector having T overhang. Presence of inserts in the recombinant plasmids was confirmed by digestion with restriction enzyme ApaI. The amplification of full length genome and its cloning will help in molecular characterization of the virus which in turn will help in developing better strategy to control the disease.