Details:

Alternative Title:
J Virol Methods
Personal Author:
Bousse, Tatiana ; Shore, David A. ; Goldsmith, Cynthia S. ; Hossain, M. Jaber ; Jang, Yunho ; ... More +
Bousse, Tatiana ; Shore, David A. ; Goldsmith, Cynthia S. ; Hossain, M. Jaber ; Jang, Yunho ; Davis, Charles T. ; Donis, Ruben O. ; Stevens, James Less -
Description:
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 had 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.
Subject:
[+]
Animals
Article
Cell Line
Chick Embryo
Dogs
Field Flow Fractionation
Fractionation, Field Flow
Influenza A Virus
Influenza Virus
Light
Multiangle Light Scattering
Reproducibility Of Results
Vaccine
Viral Load

Source:
J Virol Methods. 193(2):589-596.
Pubmed ID:
23916678
Pubmed Central ID:
PMC5695032
Document Type:
Journal Article
Funding:
CC999999/Intramural CDC HHS/United States
Collection(s):
CDC Public Access
Main Document Checksum:
[+]
urn:sha256:c3773f9ef517d166d64e5d522426653ca787eebde211657c7253e12199ba1215
File Type:

Supporting Files

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