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Production of High-Titer Infectious Influenza Pseudotyped Particles with Envelope Glycoproteins from Highly Pathogenic H5N1 and Avian H7N9 Viruses
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Imunologia e Infecção
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JoVE Journal Imunologia e Infecção
Production of High-Titer Infectious Influenza Pseudotyped Particles with Envelope Glycoproteins from Highly Pathogenic H5N1 and Avian H7N9 Viruses

Production of High-Titer Infectious Influenza Pseudotyped Particles with Envelope Glycoproteins from Highly Pathogenic H5N1 and Avian H7N9 Viruses

DOI:
10.3791/60663-v

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08:10 min

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January 15, 2020

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, , , , , ,
1Central Laboratory,Hang Zhou Red Cross Hospital, 2Clinical Laboratory,Yuhang District Maternity and Child Health Care Hospital

Capítulos

  • 00:04Título
  • 00:39Four-plasmid Cotransfection Mediated by Lipofection
  • 02:15Susceptible Cell Seeding and Pseudotyped Viral Particle Quantification
  • 04:01Infectivity Assay
  • 05:34Results: Infectivity Assay of Pseudotyped Particles
  • 06:58Conclusion

Summary

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This protocol describes an experimental process to produce high-titer infectious viral pseudotyped particles (pp) with envelope glycoproteins from two influenza A strains and how to determine their infectivity. This protocol is highly adaptable to develop pps of any other type of enveloped viruses with different envelope glycoproteins.

Tags

Keywords: InfluenzaPseudotyped ParticlesViral Envelope GlycoproteinsH5N1H7N9BSL-2TransfectionQRT-PCRVirus ReassortmentCell CultureIncubationHarvestQuantificationBenzonase Nuclease

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