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One-step RT-PCR for the detection of infectious bursal disease virus in clinical samples
R S Kataria1, A K Tiwari, T Nanthakumar
1National Biotechnology Centre, Indian Veterinary Research Institute, Izatnagar, UP.
Abstract:
A single-tube, non-interrupted, one-step RT-PCR has been standardized to amplify the hypervariable region of the VP2 gene sequence of infectious bursal disease virus (IBDV). The technique standardized on purified viral RNA was successfully applied to the detection of the virus directly in clinical samples. The amplified products were confirmed to be IBDV specific by their size in ethidium bromide-stained agarose gel, nested PCR and restriction enzyme digestion. Digestion of the amplicons with StyI restriction enzyme also differentiated classical virus from six very virulent field isolates. The sensitivity of the one-step RT-PCR was found to be 0.2 pg of viral RNA.