tetano
Editor, Senior Moderator
J Clin Microbiol. 2019 Sep 25. pii: JCM.00602-19. doi: 10.1128/JCM.00602-19. [Epub ahead of print]
[h=1]A cheap and accurate RT-PCR-melting temperature assay for real time Influenza B lineages discrimination.[/h] Motta FC[SUP]1[/SUP], Born PS[SUP]2[/SUP], Resende PC[SUP]2[/SUP], Brown D[SUP]2[/SUP], Siqueira MM[SUP]2[/SUP].
[h=3]Author information[/h] 1 WHO/NIC, Respiratory Viruses and Measles Laboratory, Oswaldo Cruz Institute, FIOCRUZ, Rio de Janeiro, Brazil fcm@ioc.fiocruz.br. 2 WHO/NIC, Respiratory Viruses and Measles Laboratory, Oswaldo Cruz Institute, FIOCRUZ, Rio de Janeiro, Brazil.
[h=3]Abstract[/h] In this work, we describe a SYBR-Green one-step RT-PCR protocol coupled with a melting temperature analysis (RT-PCR-Tm), which allows the discrimination of influenza B lineages Yamagata and Victoria. The assay is performed using a regular real time thermocycler and is based on differences in melting temperature (Tm) of a 131pb amplicon, obtained from a conserved region of hemagglutinin (HA) gene. A total of 410 samples collected during the 2004, 2008 and 2010-2017 influenza seasons in Brazil were tested and the lineages were correctly characterized using their melting profile. The temperature range is significantly different between both lineages throughout the time (Mann-Whitney test p<0.0001, CI 95%) and the Tm is not affected by viral load (Spearman Correlation test r = 0.287, p= 2.245x10[SUP]-9[/SUP]). The simplicity and cost-effectiveness of this protocol make it an option for influenza B lineage surveillance worldwide.
Copyright ? 2019 Motta et al.
PMID: 31554673 DOI: 10.1128/JCM.00602-19
Free full text
[h=1]A cheap and accurate RT-PCR-melting temperature assay for real time Influenza B lineages discrimination.[/h] Motta FC[SUP]1[/SUP], Born PS[SUP]2[/SUP], Resende PC[SUP]2[/SUP], Brown D[SUP]2[/SUP], Siqueira MM[SUP]2[/SUP].
[h=3]Author information[/h] 1 WHO/NIC, Respiratory Viruses and Measles Laboratory, Oswaldo Cruz Institute, FIOCRUZ, Rio de Janeiro, Brazil fcm@ioc.fiocruz.br. 2 WHO/NIC, Respiratory Viruses and Measles Laboratory, Oswaldo Cruz Institute, FIOCRUZ, Rio de Janeiro, Brazil.
[h=3]Abstract[/h] In this work, we describe a SYBR-Green one-step RT-PCR protocol coupled with a melting temperature analysis (RT-PCR-Tm), which allows the discrimination of influenza B lineages Yamagata and Victoria. The assay is performed using a regular real time thermocycler and is based on differences in melting temperature (Tm) of a 131pb amplicon, obtained from a conserved region of hemagglutinin (HA) gene. A total of 410 samples collected during the 2004, 2008 and 2010-2017 influenza seasons in Brazil were tested and the lineages were correctly characterized using their melting profile. The temperature range is significantly different between both lineages throughout the time (Mann-Whitney test p<0.0001, CI 95%) and the Tm is not affected by viral load (Spearman Correlation test r = 0.287, p= 2.245x10[SUP]-9[/SUP]). The simplicity and cost-effectiveness of this protocol make it an option for influenza B lineage surveillance worldwide.
Copyright ? 2019 Motta et al.
PMID: 31554673 DOI: 10.1128/JCM.00602-19
Free full text