Generating recombinant virus as a seed strain for producing A/H5N1 Avian influenza vaccine by reverse genetics

  • Nguyễn Thị Thu Hằng
  • Nguyễn Hùng Chí
  • Chu Hoàng Hà
  • Nguyễn Trung Nam
Keywords: 293T, reverse genetics, recombinant, vaccine, A/H5N1 Avian influenza virus

Abstract

Reverse genetics was applied to generate recombinant A/H5N1, clade 1.1 avian influenza virus (rg-A/H5N1 clade 1.1) as a candidate virus for producing vaccine against avian influenza viruses. Recombinant viruses were recovered at the highest titer using 8-plasmid DNA transfection
system (6:2 plasmids) containing six backborn gene segments from A/PuertoRico/8/34(H1N1) virus and two H5 (the multi-basic site was removed) and N1 antigen gene segments of A/duck/Vietnam/
ST0970/2009(H5N1) (clade 1.1) virus into 293T cells under stimulation of lipofectamine-2000 reagents for 30 minutes. The vaccine candidate strain was confirmed as a low pathogenicity avian influenza virus (LPAIV), presented genetic stability of the H5 and N1 antigen genes and grew
at high yields in embryonated chicken eggs. Inactivated oil-emulsion vaccine producing from rg-A/ H5N1, clade 1.1 virus presented ability in inducing immunogenicity to produce specific antibodies in 3-days-old pathogen-free chickens. 100% of vaccinated chickens (1024 HAU) presented HI (log2) antibody titers ≥ 4.0 after 2 and 4 weeks of vaccination

điểm /   đánh giá
Published
2021-05-17
Issue
No. 1 (2020)
Section
Nghiên cứu khoa học