The aim of this study was to improve flavivirus field monitoring in Brazil using a reliable probe-based RT-qPCR assay. Standard flavivirus strains were employed to evaluate the performance of the assay, and its applicability was evaluated using 235 stored pools of Culicidae samples collected between 1993 and 1997 and in 2016. Flavivirus species were identified by sequencing. Sixteen (6.8%) samples tested positive: Ilheus virus, Iguape virus, and Saint Louis encephalitis virus were identified in historical specimens from 1993-1994, while insect-specific flaviviruses were detected in the samples from 2016. This approach was demonstrated to be accurate for flavivirus detection and characterization, and it can be successfully applied for vector surveillance and for monitoring and discovery of insect specific flaviviruses.
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We thank the staff of the Vector-Borne Diseases Laboratory of Adolfo Lutz Institute, Akemi Suzuki, Luis Eloy Pereira, Iray Maria Rocco, Terezinha Lisieux Moraes Coimbra and Renato Pereira de Sousa, for laboratory assistance. We thank the staff of the Cellular Cultures Laboratory of Adolfo Lutz Institute, Aurea Silveira Cruz Garçon and Ana Cristina Scarparo de Miranda, for supplying cell lines. We would also like to thank Antonio Erculiani Junior for technical assistance, and the Superintendência de Controle de Endemias-SUCEN for assistance in mosquito collection and identification.
This work was also supported by FAPESP #2012/23645-4 (Dr. Paulo Cesar Mayorca) and FAPESP #2013/21719–3 (Dr. Maurício Lacerda Nogueira).
Prior approval was granted by the Ethics Committee for the Use of Animals, School of Veterinary Medicine and Animal Science, University of São Paulo, São Paulo, Brazil (CEUA no. 5400050214).
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Cunha, M.S., Luchs, A., dos Santos, F.C.P. et al. Applying a pan-flavivirus RT-qPCR assay in Brazilian public health surveillance. Arch Virol 165, 1863–1868 (2020). https://doi.org/10.1007/s00705-020-04680-w