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J Clin Virol. 2013 May 18. pii: S1386-6532(13)00158-3. doi: 10.1016/j.jcv.2013.04.018. [Epub ahead of print]
The use of pyrosequencer-generated sequence-signatures to identify the influenza B-lineage and the subclade of the B/Yamataga-lineage viruses from currently circulating human influenza B viruses.
Deng YM, Iannello P, Caldwell N, Jelley L, Komadina N, Baas C, Kelso A, Barr IG.
Source
WHO Collaborating Centre for Reference and Research on Influenza, 10 Wreckyn Street, North Melbourne, VIC 3051, Australia.
Abstract
BACKGROUND:
Influenza B viruses belong to two antigenically and genetically distinct lineages which co-circulate in varying proportions in many countries.
OBJECTIVE:
To develop simple, rapid, accurate and robust methods to detect and differentiate currently circulating B-lineage viruses in respiratory samples and virus isolates.
STUDY DESIGN:
Haemagglutinin (HA) gene sequences from more than 6300 influenza B strains were analysed to identify signature sequences that could be used to distinguish between B-lineages and sublineages.
RESULTS:
Pyrosequencing and a real time PCR assays were developed to detect the major B-lineages (B/Victoria/2/87 or B/Yamagata/16/88) and pyrosequencing for a unique mutation was used to further differentiate the B/Yamagata viruses into two currently co-circulating subgroups. More than 300 influenza virus-containing samples, including original specimens, cell and egg grown viruses, were tested with a 100% accuracy. Furthermore, when the same PCR primers were used in an rRT-PCR assay, the two lineages could be differentiated by their distinct ranges of melting temperature with an overall accuracy of 99% for 158 samples tested.
CONCLUSIONS:
These new pyrosequencing and rRT-PCR methods have the potential to aid the rapid identification of influenza B-lineages for surveillance purposes and to increase the available data for bi-annual selection of viruses for updating influenza vaccines.
Copyright ? 2013 Elsevier B.V. All rights reserved.
PMID:
23692962
[PubMed - as supplied by publisher]
The use of pyrosequencer-generated sequence-signatures to identify the influenza B-lineage and the subclade of the B/Yamataga-lineage viruses from currently circulating human influenza B viruses.
Deng YM, Iannello P, Caldwell N, Jelley L, Komadina N, Baas C, Kelso A, Barr IG.
Source
WHO Collaborating Centre for Reference and Research on Influenza, 10 Wreckyn Street, North Melbourne, VIC 3051, Australia.
Abstract
BACKGROUND:
Influenza B viruses belong to two antigenically and genetically distinct lineages which co-circulate in varying proportions in many countries.
OBJECTIVE:
To develop simple, rapid, accurate and robust methods to detect and differentiate currently circulating B-lineage viruses in respiratory samples and virus isolates.
STUDY DESIGN:
Haemagglutinin (HA) gene sequences from more than 6300 influenza B strains were analysed to identify signature sequences that could be used to distinguish between B-lineages and sublineages.
RESULTS:
Pyrosequencing and a real time PCR assays were developed to detect the major B-lineages (B/Victoria/2/87 or B/Yamagata/16/88) and pyrosequencing for a unique mutation was used to further differentiate the B/Yamagata viruses into two currently co-circulating subgroups. More than 300 influenza virus-containing samples, including original specimens, cell and egg grown viruses, were tested with a 100% accuracy. Furthermore, when the same PCR primers were used in an rRT-PCR assay, the two lineages could be differentiated by their distinct ranges of melting temperature with an overall accuracy of 99% for 158 samples tested.
CONCLUSIONS:
These new pyrosequencing and rRT-PCR methods have the potential to aid the rapid identification of influenza B-lineages for surveillance purposes and to increase the available data for bi-annual selection of viruses for updating influenza vaccines.
Copyright ? 2013 Elsevier B.V. All rights reserved.
PMID:
23692962
[PubMed - as supplied by publisher]
