Differentiation of infectious bursal disease viruses by restriction enzyme analysis of RT-PCR amplified VP1 gene sequence.
In order to differentiate infectious bursal disease virus (IBDV) isolates/strains, a quick method of RT-PCR followed by restriction enzyme analysis of VP1 gene sequence is being reported for the first time. A 480 bp fragment, comprising one of the RNA dependent RNA polymerase motifs of VP1 gene sequence of an Indian classical virus, an attenuated vaccine strain, Georgia and two Indian field isolates, genetically similar to reported very virulent strains of IBDV, was amplified by RT-PCR. Restriction enzyme digestion of PCR products with Taq1 enzyme generated distinct profile for field isolates, different from the classical and attenuated viruses, whereas restriction profile with BstNI restriction enzyme was similar in all the viruses, irrespective of the pathotype. Therefore, the present results suggest that Taq1 digestion can be taken up for the differentiation of field isolates from the classical and vaccine strains. The sequence analysis of VPI gene of reported very virulent IBD viruses from Europe and Japan, using 'MapDraw' programme of Lasergene software, revealed similar restriction enzyme profile as in Indian field isolates.
Tiwari
A K
AK
National Biotechnology Centre, Indian Veterinary Research Institute, Izatnagar 243 122, UP, India. aktiwari@ivri.up.nic.in
Kataria
R S
RS
Indervesh
Prasad
N
N
Gupta
R
R
D000818:Animals; D018175:Birnaviridae Infections / Q000662:veterinary / Q000821:virology; D002645:Chickens; D007243:Infectious bursal disease virus / Q000145:classification* / Q000235:genetics / Q000276:immunology; D011201:Poultry Diseases / Q000821:virology; D015183:Restriction Mapping / Q000379:methods / Q000662:veterinary*; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000379:methods / Q000662:veterinary*; D014613:Vaccines, Attenuated / Q000235:genetics; D015678:Viral Structural Proteins / Q000235:genetics*; D014765:Viral Vaccines / Q000235:genetics
C107669:VP1 protein, infectious bursal disease virus; D014613:Vaccines, Attenuated; D015678:Viral Structural Proteins; D014765:Viral Vaccines