02191naa a2200289 a 450000100080000000500110000800800410001902400400006010000140010024501270011426000090024152013590025065000220160965000100163165000110164165300250165265300220167765300200169965300220171965300150174165300140175665300220177070000130179270000180180570000180182377300600184121138272019-11-01       2019    bl uuuu     u00u1 u    #d7 a10.1016/j.jviromet.2019.04.0052DOI1 aHAACH, V.  aOne-step multiplex RT-qPCR for the detection and subtyping of influenza A virus in swine in Brazil.h[electronic resource]  c2019  aAbstract: 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 ubtypes 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.  aInfluenza A virus  aSwine  aSuíno  aDiagnóstico rápido  aMultiplex RT-qPCR  aRapid diagnosis  aRT-qPCR multiplex  aSubtipagem  aSubtyping  aVírus da gripe A1 aGAVA, D.1 aCANTAO, M. E.1 aFRANCO, A. C.  tJournal of Virological Methodsgv. 269, p. 43-48, 2019.