[Objective] The aim of this paper was to study the genotype, pathogenicity and nucleotide difference between Newcastle disease virus(NDV) isolates and traditional NDV vaccine strain(La Sota). [Method] A suspected NDV ...[Objective] The aim of this paper was to study the genotype, pathogenicity and nucleotide difference between Newcastle disease virus(NDV) isolates and traditional NDV vaccine strain(La Sota). [Method] A suspected NDV strain was isolated from a chicken farm. The isolate was preliminarily determined by HA and HI tests. A pair of primers was designed based on the partial sequence of NDV F gene published in GenBank(accession No. JF950510.1). F gene was amplified by RT-PCR, cloned and sequenced. The sequencing result was compared with the F gene sequences published in GenBank, and the phylogenetic tree was constructed to analyze the genotypes. The pathogenicity of the virus was determined by mean death time(MDT) of chicken embryos, intracerebral pathogenicity index(ICPI) of one-day-old chicks and intravenous pathogenicity index(IVPI) of six-week-old chickens, respectively. Based on the NDV genome sequence published in GenBank(accession No. JF950510.1), nine pairs of primers were designed to amplify the genome sequence of the isolate, and its structure was analyzed. [Result] The length of F gene was about 500 bp, and a NDV strain of genotype VII was isolated. The MDT, ICPI and IVPI were 52.8 h, 1.675 and 2.46, respectively, indicating the isolate was a virulent strain. The whole genome sequence analysis results showed that the full genome length of the isolate was 15 192 bp, which had 6 more nu-cleotides than that of La Sota strain, and the homology between the two strains was 82.8%. [Conclusion] A virulent NDV strain of genotype Ⅶ was isolated, with low homology to La Sota strain in nucleotide sequence.展开更多
基金Supported by Science and Technology Innovation Leading Talent of Qingdao City(16-8-3-14-zhc)
文摘[Objective] The aim of this paper was to study the genotype, pathogenicity and nucleotide difference between Newcastle disease virus(NDV) isolates and traditional NDV vaccine strain(La Sota). [Method] A suspected NDV strain was isolated from a chicken farm. The isolate was preliminarily determined by HA and HI tests. A pair of primers was designed based on the partial sequence of NDV F gene published in GenBank(accession No. JF950510.1). F gene was amplified by RT-PCR, cloned and sequenced. The sequencing result was compared with the F gene sequences published in GenBank, and the phylogenetic tree was constructed to analyze the genotypes. The pathogenicity of the virus was determined by mean death time(MDT) of chicken embryos, intracerebral pathogenicity index(ICPI) of one-day-old chicks and intravenous pathogenicity index(IVPI) of six-week-old chickens, respectively. Based on the NDV genome sequence published in GenBank(accession No. JF950510.1), nine pairs of primers were designed to amplify the genome sequence of the isolate, and its structure was analyzed. [Result] The length of F gene was about 500 bp, and a NDV strain of genotype VII was isolated. The MDT, ICPI and IVPI were 52.8 h, 1.675 and 2.46, respectively, indicating the isolate was a virulent strain. The whole genome sequence analysis results showed that the full genome length of the isolate was 15 192 bp, which had 6 more nu-cleotides than that of La Sota strain, and the homology between the two strains was 82.8%. [Conclusion] A virulent NDV strain of genotype Ⅶ was isolated, with low homology to La Sota strain in nucleotide sequence.