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KMID : 0381120140360050633
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Genes and Genomics
2014 Volume.36 No. 5 p.633 ~ p.641
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Analysis of genome integrity of influenza virus in formaldehyde-inactivated split vaccines
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Lee Mi-Kyung
Hwang Jae-Seon
Lee Byung-Uk
Kim Hyung-Joo
Han Jung-Soo
Lee Im-Soon
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Abstract
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Although inactivated split-virus influenza vaccines are widely used to render protection against the viral infection, information regarding the effects of the vaccine preparation on the integrity of the viral genomes is currently very limited. In the present study, to gain insight into the physical and genetic alterations of RNA viral genomes in the course of the vaccine preparation, we analyzed the influenza virus A H1N1 hemagglutinin (HA) genome segment in a conventional split-virus influenza vaccine inactivated by formaldehyde, which encodes the major surface protein of the virus. The vaccine solutions were treated with proteinase K prior to RNA purification, and viral genomic RNA was successfully retrieved up to almost eightfold higher than samples without the treatment. Despite the high yield, no full-length bands of the influenza viral genomes were identifiable upon electrophoresis due to severe degradation. Interestingly, however, we were able to amplify a variety of genomic regions including a fragment covering the full-length HA gene using cDNA. Furthermore, sequencing of a series of the fragments for confirmation revealed that the rate of base alteration of the gene in the vaccine was approximately 0.1 %, which is comparable to the spontaneous error rate occurring during PCR. We next constructed a GST expression vector carrying the full-length HA fragment cloned from the vaccine and confirmed that the correct size of the expected GST-fused HA protein was expressed. Taken together, these results demonstrate that a full-length HA RNA genome fragment with comparatively intact sequence information may exist in the inactivated split-virus vaccine. Therefore, these experimental findings on the properties of the HA RNA in the influenza vaccine, may contribute to cautious use of the vaccine in a variety of research areas and protocols.
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KEYWORD
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Influenza virus, H1N1, Hemagglutinin, Inactivated vaccine, Formaldehyde
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