tetano
Editor, Senior Moderator
J Virol Methods. 2019 Apr 5. pii: S0166-0934(19)30055-2. doi: 10.1016/j.jviromet.2019.04.005. [Epub ahead of print]
[h=1]One-step multiplex RT-qPCR for the detection and subtyping of influenza A virus in swine in Brazil.[/h] Haach V[SUP]1[/SUP], Gava D[SUP]2[/SUP], Eg?dio Cant?o M[SUP]2[/SUP], Franco AC[SUP]1[/SUP], Schaefer R[SUP]3[/SUP].
[h=3]Author information[/h]
[h=3]Abstract[/h] Pandemic H1N1, human-like H1N2 and H3N2 influenza A (IAV) viruses are co-circulating in swine herds in Brazil. The genetic analysis of the Brazilian IAVs has shown that they are genetically distinct from viruses found in swine in other countries; therefore, an update of the diagnostic assays for IAV detection and subtyping is needed. This study describes the development and validation of a TaqMan based - one-step multiplex RT-qPCR to discriminate the hemagglutinin and neuraminidase genes of the three major IAV subtypes circulating in pigs in Brazil. The RT-qPCR assays presented 100% (95.7-100, CI 95%) of diagnostic sensitivity in the analysis of 85 IAVs, previously characterized by sequencing. The limits of detection ranged from 5.09 ? 101 to 5.09 ? 103 viral RNA copies/μL. For the analytical specificity, 73 pig samples collected during 2017 and 2018 were analyzed, resulting in the identification of the subtype in 74.0% (62.9-82.7, CI 95%) of samples. From these, 46.3% were H3N2, 33.3% were H1N1, 11.1% were H1N2 and 3.7% were HxN1. Mixed viral infections (3.7%) and reassortant viruses (1.9%) were also detected by the test. This multiplex RT-qPCR assay provides a fast and specific diagnostic tool for identification of different subtypes and lineages of IAV in pigs, contributing to the monitoring of influenza in swine.
Copyright ? 2019. Published by Elsevier B.V.
[h=4]KEYWORDS:[/h] Influenza A virus; Multiplex RT-qPCR; Rapid diagnosis; Subtyping; Swine
PMID: 30959063 DOI: 10.1016/j.jviromet.2019.04.005
[h=1]One-step multiplex RT-qPCR for the detection and subtyping of influenza A virus in swine in Brazil.[/h] Haach V[SUP]1[/SUP], Gava D[SUP]2[/SUP], Eg?dio Cant?o M[SUP]2[/SUP], Franco AC[SUP]1[/SUP], Schaefer R[SUP]3[/SUP].
[h=3]Author information[/h]
[h=3]Abstract[/h] Pandemic H1N1, human-like H1N2 and H3N2 influenza A (IAV) viruses are co-circulating in swine herds in Brazil. The genetic analysis of the Brazilian IAVs has shown that they are genetically distinct from viruses found in swine in other countries; therefore, an update of the diagnostic assays for IAV detection and subtyping is needed. This study describes the development and validation of a TaqMan based - one-step multiplex RT-qPCR to discriminate the hemagglutinin and neuraminidase genes of the three major IAV subtypes circulating in pigs in Brazil. The RT-qPCR assays presented 100% (95.7-100, CI 95%) of diagnostic sensitivity in the analysis of 85 IAVs, previously characterized by sequencing. The limits of detection ranged from 5.09 ? 101 to 5.09 ? 103 viral RNA copies/μL. For the analytical specificity, 73 pig samples collected during 2017 and 2018 were analyzed, resulting in the identification of the subtype in 74.0% (62.9-82.7, CI 95%) of samples. From these, 46.3% were H3N2, 33.3% were H1N1, 11.1% were H1N2 and 3.7% were HxN1. Mixed viral infections (3.7%) and reassortant viruses (1.9%) were also detected by the test. This multiplex RT-qPCR assay provides a fast and specific diagnostic tool for identification of different subtypes and lineages of IAV in pigs, contributing to the monitoring of influenza in swine.
Copyright ? 2019. Published by Elsevier B.V.
[h=4]KEYWORDS:[/h] Influenza A virus; Multiplex RT-qPCR; Rapid diagnosis; Subtyping; Swine
PMID: 30959063 DOI: 10.1016/j.jviromet.2019.04.005