Pathotyping of Newcastle Disease Viruses by RT-PCR and Restriction Enzyme Analysis |
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| Authors: | Nanthakumar T Kataria RS Tiwari AK Butchaiah G Kataria JM |
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| Institution: | (1) National Biotechnology Centre, Indian Veterinary Research Institute, Izatnagar, U.P. 243 122, India |
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| Abstract: | The technique of RT-PCR and restriction enzyme analysis was standardized to detect and differentiate Newcastle disease viruses. Digestion of RT-PCR-amplified, F gene sequences encoding for the cleavage activation sites of fusion protein with restriction enzymes AluI, BglI, HaeIII, HinfI, HhaI, RsaI, StyI and TaqI was carried out in order to characterize Newcastle disease viruses of varying pathogenicity. Restriction enzyme digestion of the amplicons by BglI and HhaI could group eight viruses, both field isolates and known vaccine strains, into lentogenic, mesogenic and velogenic pathotypes. By employing this technique directly on a clinical sample, Newcastle disease virus of the lentogenic pathotype could be detected. |
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| Keywords: | amplicons classification diagnosis Newcastle disease pathotype polymerase chain reaction restriction enzymes |
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