Construction of Indonesian-Strain Avian Flu Virus Seed Vaccine Using Low Pathogenic Hemagglutinin Gene and Neuraminidase Pr8 Gene through Reverse Genetics

Reviany Vibriaanita Nidom, Muh Y Alamudi, Sahrir Sillehu, Setyarina Indrasari, Rahmalia D Suindarti, Ema Qurnianingsih, Yoes P Dachlan, Aryati, Ahmad Syahrani, Kadek Rachmawati, Kuncoro P Santoso and Chairul A Nidom*

Avian Influenza H5N1 has been spreading in Indonesia since 2003. The Avian influenza virus H5N1 actually infects various animals, including birds, mammalian and others. Since 2005, avian influenza virus has infected humans. According to data, there are 132 persons suspected of contracting this virus, and this was one of the highest cases of avian influenza H5N1 outbreak in the world. One of the means of protection against infection is vaccination. Until now there is no H5N1 influenza vaccine extant, because the virus has a high level of pathogenicity, which makes it difficult to obtain a high yield of seed vaccine.

In this research, we constructed H5N1 virus with low pathogenic Hemagglutinin gene and neuraminidase gene from PR8 (H1N1) using reverse genetic method. The new virus then inoculated in embryonated chicken eggs. We tested the H5N1 virus reverse genetic using molecular methods such as PCR and sequencing; and we used EID50 and Intravenous Pathogenicity Index for the pathogenicity test. The new H5N1 virus reverse genetic was also tested in mice to observe the antibody’s titer and challenge.

The result, we found that the low pathogenic H5N1 virus reverse genetic was successfully created. Neuraminidase from PR8 played important role in reducing pathogens in chickens and mice. From molecular data we note that no amino acids were changed, the pathogenicity of H5N1 virus reverse genetic was low, with an Intravenous Pathogenicity Index of 0 and the highest titer was 256 GMT and the highest survival rate was 85.71%.

Keywords: Avian infl uenza; H5N1; Reverse genetic; Low pathogenic

Published on: Aug 17, 2017 Pages: 5-11

Full Text PDF Full Text HTML DOI: 10.17352/jvi.000021
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