tetano
Editor, Senior Moderator
J Clin Microbiol. 2013 Nov 13. [Epub ahead of print]
Incorporation of a Proteotyping Approach using Mass Spectrometry for the Surveillance of the Influenza Virus in Cell Cultured Strains.
Fernandes ND, Downard KM.
Source
School of Molecular Bioscience, University of Sydney, Sydney, Australia.
Abstract
The reemergence of deadly pandemic influenza virus strains has necessitated the development of improved methods for the rapid detection and subtyping of influenza viruses that will enable more strains to be characterized at the molecular level. Representative circulating strains of human influenza viruses from primary clinical specimens were grown in cell culture, purified through polyethylene glycol precipitation, proteolytically digested with an endoproteinase, and analysed and identified by high resolution mass spectrometry using unique signature peptides that are characteristic of type A H1N1, H3N2 and type B influenza viruses. This proteotyping approach enabled circulating strains of type A influenza virus to be typed and subtyped, co-circulating seasonal and pandemic H1N1 viruses to be differentiated and the lineage of type B viruses to be determined through single ion detection by high resolution mass spectrometry. Results were obtained using virus titres comparable to those used in RT-PCR from clinical specimens grown in cell culture. The methodology represents a more rapid and direct approach over RT-PCR that can be integrated into existing procedures currently used for the surveillance of emerging pandemic and seasonal influenza viruses.
PMID:
24226917
[PubMed - as supplied by publisher]
http://www.ncbi.nlm.nih.gov/pubmed/24226917
Incorporation of a Proteotyping Approach using Mass Spectrometry for the Surveillance of the Influenza Virus in Cell Cultured Strains.
Fernandes ND, Downard KM.
Source
School of Molecular Bioscience, University of Sydney, Sydney, Australia.
Abstract
The reemergence of deadly pandemic influenza virus strains has necessitated the development of improved methods for the rapid detection and subtyping of influenza viruses that will enable more strains to be characterized at the molecular level. Representative circulating strains of human influenza viruses from primary clinical specimens were grown in cell culture, purified through polyethylene glycol precipitation, proteolytically digested with an endoproteinase, and analysed and identified by high resolution mass spectrometry using unique signature peptides that are characteristic of type A H1N1, H3N2 and type B influenza viruses. This proteotyping approach enabled circulating strains of type A influenza virus to be typed and subtyped, co-circulating seasonal and pandemic H1N1 viruses to be differentiated and the lineage of type B viruses to be determined through single ion detection by high resolution mass spectrometry. Results were obtained using virus titres comparable to those used in RT-PCR from clinical specimens grown in cell culture. The methodology represents a more rapid and direct approach over RT-PCR that can be integrated into existing procedures currently used for the surveillance of emerging pandemic and seasonal influenza viruses.
PMID:
24226917
[PubMed - as supplied by publisher]
http://www.ncbi.nlm.nih.gov/pubmed/24226917