Gene VII type Newcastle disease virus attenuated strain, vaccine composition and application of vaccine composition
A vaccine composition and a technology for Newcastle disease virus, which are applied in the field of attenuated strains of genotype VII Newcastle disease virus, can solve problems such as protection defects, achieve good immunogenicity, reduce the rate of virulent infection and excretion, and have a good immune effect. Effect
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[0015] As a preferred embodiment of the present invention, the vaccine composition includes an immune amount of antigens of gene type VII attenuated Newcastle disease vaccine strain N7a strain or 1-60 passage cultures thereof. More preferably, the vaccine composition includes an immune amount of the gene type VII attenuated Newcastle disease vaccine strain N7a strain or its 1-55 generation cultured antigen; or an immune amount of the gene type VII Newcastle disease attenuated vaccine strain N7a strain Or the antigen of its 1-50th generation culture; or the antigen of gene Ⅶ type Ⅶ attenuated Newcastle disease vaccine strain N7a strain or its 1-45 generation culture including immune amount; or the gene Ⅶ type Ⅶ Newcastle disease attenuated virus The antigen of vaccine strain N7a strain or its 1-40th generation culture; or the antigen of gene VII type Newcastle disease attenuated vaccine strain N7a strain or its 1-35th generation culture including immunization amount; or the gene...
Embodiment 1
[0046] Example 1 Construction of Gene Type VII Newcastle Disease Attenuated Vaccine Strain
[0047] The whole genome cDNA clone (transcription vector) was constructed by segmental cloning method, and the F gene cleavage site 112R-R-Q-K-R-F117 in the genome of the isolated gene VII type virulent Newcastle disease strain PLK-N-06 strain was isolated by Overlap PCR Mutate to the attenuated strain characteristic 112G-R-Q-G-R-L117, and clone NP, P and L into the eukaryotic expression vector pCI-neo to be the helper plasmid for NDV rescue. The rescue process of NDV is to co-transfect BHK-21 cells with the whole genome cDNA clone and the helper plasmid containing the genes NP, P and L, and under the action of the relevant enzymes provided by the helper plasmid, the cDNA clone is transcribed to produce genome positive single-stranded RNA , the polymerase protein expressed by the helper plasmid wraps the positive-sense single-stranded RNA of the genome to form ribonucleoprotein complex...
Embodiment 2
[0049] The biological characteristics of embodiment 2 N7a strain Newcastle disease virus attenuated vaccine strain
[0050] The hemagglutination value (HA), the mean lethal time (MDT) of chicken embryos, the pathogenicity index (ICPI) and the median lethal dose (ELD) of 1-day-old chicks were determined for attenuated Newcastle disease virus vaccine strain N7a strain. 50 ). The specific method is as follows.
[0051] The hemagglutination value was determined according to the current "Chinese Veterinary Pharmacopoeia".
[0052] Chicken embryo mean lethal time (MDT) determination method:
[0053] (1) The freshly infected allantoic fluid was serially diluted 10 times with sterilized normal saline to 10 -6 ~10 -9 ;
[0054] (2) Inoculate 10 9-11-day-old SPF chicken embryos into the allantoic cavity of each dilution, each inoculate 0.1ml, and culture at 37°C;
[0055] (3) illuminate eggs 4 times a day, observe continuously for 7 days, and record the death time of each chicken em...
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