Virus Characterization and Sequence of the Fusion Protein Gene Cleavage Site of Recent Newcastle Disease Virus Field Isolates from the Southeastern United States and Puerto Rico

Abstract

Nine Newcastle disease virus (NDV) isolates obtained from Puerto Rico, Georgia, Alabama, Mississippi, and Texas were analyzed for in vivo pathogenicity, biological properties (hemagglutination of mammalian erythrocytes), and for sequence variation at the amino acid and sense RNA level of the fusion protein cleavage site. Intracerebral pathogenicity index values ranged from 0 to 0.3 and the intravenous pathogenicity index obtained for all isolates was 0. Four isolates hemagglutinated bovine erythrocytes, whereas no hemagglutination was observed using equine erythrocytes. By direct sequencing of reverse transcription polymerase chain reaction products, all the isolates had a predicted fusion cleavage sequence comparable to lentogenic NDV strains. Based on nucleotide sequence, the viruses could be grouped phylogenetically with the B1 vaccine-type virus. However, nucleotide sequences were not 100% similar to the B1 or La Sota NDV strains, indicating that minor genetic heterogeneity occurs among lentogenic field isolates of NDV.

Molecular epidemiology of Newcastle disease viruses isolated in South Korea using sequencing of the fusion protein cleavage site region and phylogenetic relationships

“…These multiple basic amino acid sequences can be recognized by ubiquitous host proteinases. Pathotype prediction and diagnosis of the virulence of NDV can be performed using the fusion protein cleavage site sequence analysis (Collins et al, 1993;Seal et al, 1995;Marin et al, 1996;Gould et al, 2001).…”

“…Various methods have been introduced for grouping and analysis of NDV including differences in pathogenicity (Alexander, 1997), antigenicity (Russell & Alexander, 1983;Alexander et al, 1987Alexander et al, , 1997, and genomes (Collins et al, 1993;Jarecki-Black & King, 1993;Seal et al, 1995Seal et al, , 1996Seal et al, , 2000Ballagi-Pordany et al, 1996;Marin et al, 1996;Kant et al, 1997;Lomniczi et al, 1998;Herczeg et al, 1999Herczeg et al, , 2001Yang et al, 1999;Nanthakumar et al, 2000;Aldous et al, 2001Aldous et al, , 2003Gould et al, 2001;Ke et al, 2001;Yu et al, 2001;Liang et al, 2002;Mase et al, 2002;Liu et al, 2003;Wehmann et al, 2003a,b). Several epidemiological studies of NDV have been carried out using molecular-based methods.…”

Differentiation of virulent and non‐virulent strains of Newcastle disease virus within 24 hours by polymerase chain reaction

“…Nucleotide sequencing of the F gene made it clear that this variable cleavability is caused by different amino acid compositions at the cleavage site (Glickman et al, 1988;Toyoda et al, 1987). Several papers have been published about differentiation of NDV strains using RT-PCR, in combination with restriction-enzyme analysis (Collins et al, 1993(Collins et al, , 1994Jestin & Cherbonnel, 1992;Jestin et al, 1994;Marin et al, 1996;Oberdorfer & Werner, 1995;Seal et al, 1995;Stauber et al, 1995). Viruses could also be differentiated by using an oligonucleotide probe, representing the sequence of virulent NDV strains at the cleavage site of the F protein.…”

Restriction Fragment Length Polymorphism Analysis of the F Gene of Newcastle Disease Viruses Isolated from Chickens and an Owl in Taiwan.

“…Molecular epidemiology has been employed to explore the sequence variation and phylogenic relationship of NDV isolates [6][7][8][9]. However, investigation of the nucleotide sequence is time-consuming when the number of samples is large, and a relatively simple way of grouping different isolates with epizootiological relatedness has been demonstrated [4].…”

“…In order to identify viruses of high and low pathogenicity in the laboratory, three pathogenicity tests, the mean death time (MDT), the intracerebral pathogenicity index (ICPI), and the intravenous pathogenicity index (IVPI), are performed [1]. These pathogenicity tests can distinguish viruses according to the degree of severity of ND caused by different isolates, but do not give much epizootiological information.Molecular epidemiology has been employed to explore the sequence variation and phylogenic relationship of NDV isolates [6][7][8][9]. However, investigation of the nucleotide sequence is time-consuming when the number of samples is large, and a relatively simple way of grouping different isolates with epizootiological relatedness has been demonstrated [4].…”