tetano
Editor, Senior Moderator
J Virol Methods. 2012 Feb 22. [Epub ahead of print]
Detection of influenza A viruses with a portable real-time PCR instrument.
M?ls? M, Koskela KA, R?nkk? E, Ikonen N, Ziegler T, Nikkari S.
Source
Centres for Military Medicine and for Biological Threat Preparedness, Tukholmankatu 8A, FI-00290 Helsinki, Finland.
Abstract
Timely identification of respiratory pathogens is essential for appropriate patient care and cohorting. In order to do rapid identification-technology near the patient we utilized the field-deployable RAZOR EX-thermocycler with a reverse transcription real-time PCR assay that detects all subtypes of influenza A virus. In addition, we developed a RT PCR assay for specific detection of influenza A(H1N1)pdm09 virus. These assays amplified segments of the matrix (M)- and the hemagglutinin (HA)-gene, respectively. Detection limits of the M-gene and the influenza A(H1N1)pdm09-specific HA-gene assays were 0.15PFU and 8.8PFU per reaction, respectively. With 18 influenza A viruses of different subtypes and influenza B, C, and 7 other respiratory viruses the RAZOR EX and standard real-time PCR assay results were in total agreement. From 104 clinical samples identical results were obtained by both PCR methods. Additional 21 clinical samples were tested under field conditions with the RAZOR EX instrument. Results were achieved in 90min, including 45min for sample preparation and they were in complete agreement with those obtained by standard real-time PCR under laboratory conditions. These methods enable highly sensitive and rapid on-site diagnostics to reliably identify patients infected with influenza A, including the influenza A(H1N1)pdm09-virus.
Copyright © 2012. Published by Elsevier B.V.
PMID:
22374102
[PubMed - as supplied by publisher]
http://www.ncbi.nlm.nih.gov/pubmed/22374102
Detection of influenza A viruses with a portable real-time PCR instrument.
M?ls? M, Koskela KA, R?nkk? E, Ikonen N, Ziegler T, Nikkari S.
Source
Centres for Military Medicine and for Biological Threat Preparedness, Tukholmankatu 8A, FI-00290 Helsinki, Finland.
Abstract
Timely identification of respiratory pathogens is essential for appropriate patient care and cohorting. In order to do rapid identification-technology near the patient we utilized the field-deployable RAZOR EX-thermocycler with a reverse transcription real-time PCR assay that detects all subtypes of influenza A virus. In addition, we developed a RT PCR assay for specific detection of influenza A(H1N1)pdm09 virus. These assays amplified segments of the matrix (M)- and the hemagglutinin (HA)-gene, respectively. Detection limits of the M-gene and the influenza A(H1N1)pdm09-specific HA-gene assays were 0.15PFU and 8.8PFU per reaction, respectively. With 18 influenza A viruses of different subtypes and influenza B, C, and 7 other respiratory viruses the RAZOR EX and standard real-time PCR assay results were in total agreement. From 104 clinical samples identical results were obtained by both PCR methods. Additional 21 clinical samples were tested under field conditions with the RAZOR EX instrument. Results were achieved in 90min, including 45min for sample preparation and they were in complete agreement with those obtained by standard real-time PCR under laboratory conditions. These methods enable highly sensitive and rapid on-site diagnostics to reliably identify patients infected with influenza A, including the influenza A(H1N1)pdm09-virus.
Copyright © 2012. Published by Elsevier B.V.
PMID:
22374102
[PubMed - as supplied by publisher]
http://www.ncbi.nlm.nih.gov/pubmed/22374102