Construction and immunogenicity of a gE/gI/TK-deleted PRV based on porcine pseudorabies virus variant
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Pseudorabies
One virus was isolated from brain organ of diseased pig which came from a pig farm in Tianjin. After inoculating the virus,the white spots could be seen in chick embryo chorioallantoic membrane, and typical cytopathogenic effect(CPE)appeared in PK-15 and BHK-21 cells. TCID50 of the virus was 10-6/mL.The virus could be neutralized by standard positive serum of pseudorabies virus. After injecting the virus, rabbits showed typical pseudorabies clinical symptom and died. By the above experiment results, the isolated virus was identified as PRV.
Pseudorabies
Chorioallantoic membrane
Virus isolation
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One Pseudorabies(Aujeszky's disease) virus isolated from infected domestic pigs originating from Wujiaqu shihezi of ×injiang.The organ samples were filtered then drained monolayers of BHK-21 cells,the typical cytopathic effect(cpe) were observed. The virus appeared typical Pseudorabies virus form:diameter was about 100 nm,circle with peplos and peplomer .The virus were neutralized with standard positive porcine sera using serum neutralization test(SNT). TCID_(50) of virus was 10-10.87/0.1mL. Rabbits showed typical syndrome of PR when they were inoculated with cells virus. PCR technique was applied for the detection PRV in cells and Amplified a fragment with desired size of 217 bp.According with virus characterisation from infected pigs,the virus was indentified as PRV.
Pseudorabies
Cytopathic effect
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One virus strain has been isolated from the samples of brain and visceral organs of a death piglet from a hogpen in Henan province.The virus was continually inoculated 5 generations onto PK15 cells and induced typical cytopathogenic effect(CPE).The typical pseudorabies virus particles could be found under electro-microscope.The isolate could be neutralized by the positive serum to pseudorabies virus.Rabbits and mice were injected with the isolate and the typical clinical syndrome could be observed.The sequence of 928 bases lying in gE gene could be amplified by the DNA extracted from the isolate.All of the evidences indicated that the isolated virus was pseudorabies virus and was named HN strain.
Pseudorabies
Strain (injury)
Alphaherpesvirinae
Virus strain
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Pseudorabies
Arterivirus
Viral culture
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The brain tissue of piglet was obtained and the 217 bp long fragment in gp50 gene was amplified by the polymerase chain reaction(PCR).Therefore,the piglet was confirmed as porcine pseudorabies virus infection.Subsequently,the porcine pseudorabies virus was isolated on BHK-21 cells after 5 generations.The cells displayed typical cell lesions and were identified as porcine pseudorabies virus by PCR.The virus infectivety of the pseudorabies virus was 108.68 TCID50/0.1 mL by Reed-Muench.Finally,the rabbits were inoculated with 107.0 TCID50/mL virus.On 48 h the injection sites showed typical itching,skin damage and other symptoms.The rabbits were dead after 72 h by the inoculation of pseudorabies virus.Consequently,the results indicated that the isolated pseudorabies virus was highly pathogenicity to susceptible animals.It would be the research foundation of the virus epidemiology,pathogenesis,vaccine and diagnositic.
Pseudorabies
Alphaherpesvirinae
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Porcine pseudorabies virus was isolated from the brain of a 2-day old pig.The Balb/c mice were inoculated with the virus and the typical PRV symptoms could be observed.Infected BHK-21 cell line shrunk,aggregated,and detached from the culture system in 48h approximately.The primers which were designed according to the sequence of PRV gE gene published in GenBank could amplify 354 bp fragment from the isolated virus.These evidences indicate that the isolated virus was PRV.
Pseudorabies
Virus isolation
Alphaherpesvirinae
Isolation
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Glycoprotein gIII of pseudorabies virus is a major antigen found in the envelopes of virus particles as well as in and on the surfaces of infected cells. It is not an essential gene product for virus growth in tissue culture. In this report, we provide evidence that, although it is not essential, the gIII protein is required for efficient virus growth and that gIII mutants are quickly outgrown by wild-type virus in mixed infections.
Pseudorabies
Alphaherpesvirinae
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Pseudorabies
Serial passage
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An isolate suspected to be pseudorabies virus was recovered from dead piglets on a pig farm in Panyu Guangdong. It was characterized by its physicochemical properties, virus neutralization test, animal inoculation, electron microscopy, PCR amplification and in vitro infectivity test. The isolate was found to be sensitive both to chloroform and ether, could be neutralized by the standard pseudorabies Fujian A strain virus antibodies, rabbits inoculated with the isolate developed localized itch and then paralysis before death, electron microscope revealed irregularly round virus particles with envelope and peplomers, PCR amplification of the virus genome yielded a fragment which co migrated with similarly amplified fragment of the Fujian A standard PRV. When titrated in Marc 145 cell culture, the virus titre attained 10 7.5 TCID 50 /mL. The isolate was identified as a pseudorabies virus on the strength of the above findings.
Pseudorabies
Infectivity
Strain (injury)
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Pseudorabies
Alphaherpesvirinae
Respiratory tract
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