tetano
Editor, Senior Moderator
J Clin Virol. 2012 Jun 4. [Epub ahead of print]
Monitoring seasonal influenza A evolution: Rapid 2009 pandemic H1N1 surveillance with an reverse transcription-polymerase chain reaction/electro-spray ionization mass spectrometry assay.
Jeng K, Massire C, Zembower TR, Deyde VM, Gubareva LV, Hsieh YH, Rothman RE, Sampath R, Penugonda S, Metzgar D, Blyn LB, Hardick J, Gaydos CA.
Source
Johns Hopkins University, Baltimore, MD, USA; Duke University, Durham, NC, USA.
Abstract
BACKGROUND:
The emergence of the pandemic H1N1 influenza strain in 2009 reinforced the need for improved influenza surveillance efforts. A previously described influenza typing assay that utilizes RT-PCR coupled to electro-spray ionization mass spectrometry (ESI-MS) played an early role in the discovery of the pandemic H1N1 influenza strain, and has potential application for monitoring viral genetic diversity in ongoing influenza surveillance efforts.
OBJECTIVES:
To determine the analytical sensitivity of RT-PCR/ESI-MS influenza typing assay for identifying the pandemic H1N1 strain and describe its ability to assess viral genetic diversity.
STUDY DESIGN:
Two sets of pandemic H1N1 samples, 190 collected between April and June of 2009, and 69 collected between October 2009 and January 2010, were processed by the RT-PCR/ESI-MS influenza typing assay, and the spectral results were compared to reference laboratory results and historical sequencing data from the Nucleotide Database of the National Center for Biotechnology Information (NCBI).
RESULTS:
Strain typing concordance with reference standard testing was 100% in both sample sets, and the assay demonstrated a significant increase in influenza genetic diversity, from 10.5% non-wildtype genotypes in early samples to 69.9% in late samples (P<0.001). An NCBI search demonstrated a similar increase, from 13.4% to 45.2% (P<0.001).
CONCLUSIONS:
This comparison of early versus late influenza samples analyzed by RT-PCR/ESI-MS demonstrates the influenza typing assay's ability as a universal influenza detection platform to provide high-fidelity pH1N1 strain identification over time, despite increasing genetic diversity in the circulating virus. The genotyping data can also be leveraged for high-throughput influenza surveillance.
Copyright ? 2012. Published by Elsevier B.V.
PMID:
22673129
[PubMed - as supplied by publisher]
http://www.ncbi.nlm.nih.gov/pubmed/22673129
Monitoring seasonal influenza A evolution: Rapid 2009 pandemic H1N1 surveillance with an reverse transcription-polymerase chain reaction/electro-spray ionization mass spectrometry assay.
Jeng K, Massire C, Zembower TR, Deyde VM, Gubareva LV, Hsieh YH, Rothman RE, Sampath R, Penugonda S, Metzgar D, Blyn LB, Hardick J, Gaydos CA.
Source
Johns Hopkins University, Baltimore, MD, USA; Duke University, Durham, NC, USA.
Abstract
BACKGROUND:
The emergence of the pandemic H1N1 influenza strain in 2009 reinforced the need for improved influenza surveillance efforts. A previously described influenza typing assay that utilizes RT-PCR coupled to electro-spray ionization mass spectrometry (ESI-MS) played an early role in the discovery of the pandemic H1N1 influenza strain, and has potential application for monitoring viral genetic diversity in ongoing influenza surveillance efforts.
OBJECTIVES:
To determine the analytical sensitivity of RT-PCR/ESI-MS influenza typing assay for identifying the pandemic H1N1 strain and describe its ability to assess viral genetic diversity.
STUDY DESIGN:
Two sets of pandemic H1N1 samples, 190 collected between April and June of 2009, and 69 collected between October 2009 and January 2010, were processed by the RT-PCR/ESI-MS influenza typing assay, and the spectral results were compared to reference laboratory results and historical sequencing data from the Nucleotide Database of the National Center for Biotechnology Information (NCBI).
RESULTS:
Strain typing concordance with reference standard testing was 100% in both sample sets, and the assay demonstrated a significant increase in influenza genetic diversity, from 10.5% non-wildtype genotypes in early samples to 69.9% in late samples (P<0.001). An NCBI search demonstrated a similar increase, from 13.4% to 45.2% (P<0.001).
CONCLUSIONS:
This comparison of early versus late influenza samples analyzed by RT-PCR/ESI-MS demonstrates the influenza typing assay's ability as a universal influenza detection platform to provide high-fidelity pH1N1 strain identification over time, despite increasing genetic diversity in the circulating virus. The genotyping data can also be leveraged for high-throughput influenza surveillance.
Copyright ? 2012. Published by Elsevier B.V.
PMID:
22673129
[PubMed - as supplied by publisher]
http://www.ncbi.nlm.nih.gov/pubmed/22673129