tetano
Editor, Senior Moderator
J Virol Methods. 2013 Jul 31. pii: S0166-0934(13)00280-2. doi: 10.1016/j.jviromet.2013.07.026. [Epub ahead of print]
Quantitation of influenza virus using field flow fractionation and multi-angle light scattering for quantifying influenza A particles.
Bousse T, Shore DA, Goldsmith CS, Hossain MJ, Jang Y, Davis CT, Donis RO, Stevens J.
Source
Influenza Division, National Center for Immunization and Respiratory Diseases, Centers for Disease Control and Prevention, Atlanta, GA, United States.
Abstract
Recent advances in instrumentation and data analysis in field flow fractionation and multi-angle light scattering (FFF-MALS) have enabled greater use of this technique to characterize and quantitate viruses. In this study, the FFF-MALS technique was applied to the characterization and quantitation of type A influenza virus particles to assess its usefulness for vaccine preparation. The use of FFF-MALS for quantitation and measurement of control particles provided data accurate to within 5% of known values, reproducible with a coefficient of variation of 1.9%. The methods, sensitivity and limit of detection were established by analyzing different volumes of purified virus, which produced a linear regression with fitting value R2 of 0.99. FFF-MALS was further applied to detect and quantitate influenza virus in the supernatant of infected MDCK cells and allantoic fluids of infected eggs. FFF fractograms of the virus present in these different fluids revealed similar distribution of monomeric and oligomeric virions. However, the monomer fraction of cell grown virus has greater size variety. Notably, β-propialactone (BPL) inactivation of influenza viruses did not influence any of the FFF-MALS measurements. Quantitation analysis by FFF-MALS was compared to infectivity assays and real-time RT-PCR (qRT-PCR) and the limitations of each assay were discussed.
Published by Elsevier B.V.
KEYWORDS:
Field flow fractionation, Influenza virus, Multiangle light scattering, Vaccine
PMID:
23916678
[PubMed - as supplied by publisher]
http://www.ncbi.nlm.nih.gov/pubmed/23916678
Quantitation of influenza virus using field flow fractionation and multi-angle light scattering for quantifying influenza A particles.
Bousse T, Shore DA, Goldsmith CS, Hossain MJ, Jang Y, Davis CT, Donis RO, Stevens J.
Source
Influenza Division, National Center for Immunization and Respiratory Diseases, Centers for Disease Control and Prevention, Atlanta, GA, United States.
Abstract
Recent advances in instrumentation and data analysis in field flow fractionation and multi-angle light scattering (FFF-MALS) have enabled greater use of this technique to characterize and quantitate viruses. In this study, the FFF-MALS technique was applied to the characterization and quantitation of type A influenza virus particles to assess its usefulness for vaccine preparation. The use of FFF-MALS for quantitation and measurement of control particles provided data accurate to within 5% of known values, reproducible with a coefficient of variation of 1.9%. The methods, sensitivity and limit of detection were established by analyzing different volumes of purified virus, which produced a linear regression with fitting value R2 of 0.99. FFF-MALS was further applied to detect and quantitate influenza virus in the supernatant of infected MDCK cells and allantoic fluids of infected eggs. FFF fractograms of the virus present in these different fluids revealed similar distribution of monomeric and oligomeric virions. However, the monomer fraction of cell grown virus has greater size variety. Notably, β-propialactone (BPL) inactivation of influenza viruses did not influence any of the FFF-MALS measurements. Quantitation analysis by FFF-MALS was compared to infectivity assays and real-time RT-PCR (qRT-PCR) and the limitations of each assay were discussed.
Published by Elsevier B.V.
KEYWORDS:
Field flow fractionation, Influenza virus, Multiangle light scattering, Vaccine
PMID:
23916678
[PubMed - as supplied by publisher]
http://www.ncbi.nlm.nih.gov/pubmed/23916678