The deletion of an extra six nucleotides in the 5′ -untranslated region of the nucleoprotein gene of Newcastle disease virus NA-1 decreases virulence

BACKGROUND: The virulent Newcastle disease virus (NDV) strain NA-1 (genotype VII) was isolated from an epizootic involving a flock of geese in Jilin Province, Northeast China, in 1999. Compared with the classical NDV strains, which have a genome size of 15,186 bp, the more recently isolated NDV strains, including that involved in the goose outbreak, have an extra six nucleotides in the 5′-untranslated region (UTR) of the nucleoprotein (NP) gene. This extra sequence, TCCCAC, is highly conserved and has been found in multiple NDV strains, including ZJ-1, WF00G, JSG0210, and NA-1. In the current study, an infectious clone from strain NA-1 was isolated and designated rNA-1. Subsequently, strain rNA-1 was mutated to delete the six-nucleotide insertion, producing strain rNA-1(−). Virulence of the recombinant virus was then assayed in chickens and geese. RESULTS: The recombinant virus rNA-1(−) showed similar biological characteristics to the parental NA-1 strain in DF-1 chicken fibroblast cells. However, the virulence of rNA-1(−) in 9-day-old embryonated chicken eggs and 1-day-old specific pathogen-free (SPF) chickens was decreased compared with the rNA-1 control. Furthermore, the virulence of the recombinant strain was slightly decreased in 1-day-old SPF chickens when compared with that in 1-day-old geese. CONCLUSION: Following deletion of six nucleotides in the 5′-UTR of the NP gene of NDV strain NA-1, the virulence of the rNA-1(−) recombinant strain was decreased in both chickens and geese. However, rNA-1(−) was more virulent in chickens than geese, as seen by the higher intracerebral pathogenicity index value.