Phylogenetic Analysis of Newcastle Disease Virus Genotypes Isolated in Japan

Abstract

We genetically analyzed field isolates of the Newcastle disease (ND) virus isolated in Japan from 1930 to 2001. The coding region of the fusion protein was amplified by reverse transcriptase PCR and directly sequenced. Phylogenetic analysis revealed the presence of viruses belonging to six of the eight known genotypes. It can be concluded from this study that ND outbreaks in Japan have been of multiple etiologies

Simple Differentiation of Avirulent and Virulent Strains of Avian Paramyxovirus Serotype-1 (Newcastle Disease Virus) by PCR and Restriction Endonuclease Analysis in Japan

“…These results demonstrated that REA of the PCR products using Hin1 I and Apa I facilitated the differentiation of avirulent and virulent field Japanese APMV-1 strains. This method was applied to other NDV strains isolated in Japan [9,10], and the same results were obtained (data not shown). This demonstrated that our identification method was performed correctly.…”

“…First, we selected representative APMV-1 strains for each genotype isolated in Japan (Table 1) [9,10]. Allantoic cavities of specific pathogen-free embryonated eggs were used for virus propagation.…”

“…Reverse transcription (RT), PCR amplification and sequencing were performed as described previously [9,10]. Briefly, after reverse transcription with SuperScript™ III (Life Technologies, Gaithersburg, MD, U.S.A.) using random 9 mers, PCR was performed to amplify cDNAs using Takara ExTaq (Takara, Tokyo, Japan).…”

Molecular and Serological Detection of Newcastle Disease Virus in Native Chickens from Selected Live Bird Markets in Batangas, Philippines

“…Extracted viral RNA were transcribed to complementary DNA (cDNA) using random hexamers and reverse transcriptase (Takara Bio-Inc,Shiga, Japan). A two-step nested RT-PCR was performed to amplify the region comprising the 3′ end of the M-gene and the 5′ end of the F-gene using SapphireAmp Fast PCR Master Mix (Takara Bio-Inc,Shiga, Japan) (25µL/50µl reaction volume), external and internal primers (0.2µM) and template (~50-100ng) as described previously (Mase et al, 2002;Umali et al, 2013;Umali et al, 2014;Umali et al, 2015). The PCR product of both first and second PCR were analysed using gel electrophoresis.…”

Newcastle disease outbreaks in Kazakhstan and Kyrgyzstan during 1998, 2000, 2001, 2003, 2004, and 2005 were caused by viruses of the genotypes VIIb and VIId

“…The sequenced fragments of the F gene were compiled and edited using a Lasergene sequence analysis software package (DNA Star software version 5.0, Madison, WI, USA). Phylogenic analysis was conducted with MEGA4: molecular evolutionary genetics analysis (MEGA) software version 4.0 to complete comparisons of the nucleotide sequences of our isolates with those of ND viruses obtained from the GenBank database [1,6,[9][10][11][12][13][14][15][16][17][18][21][22][23][24][25][26][27][28][29][30][31][32][33][34] The nucleotide regions used in the phylogenic analysis were fragments of F gene (from 47 to 374). The nucleotide sequences were deposited in GenBank and filed under the accession numbers: FJ385767-FJ385771, FJ434381-FJ434400, and FJ455441-FJ455443 (Table 1).…”