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Applying a pan-flavivirus RT-qPCR assay in Brazilian public health surveillance

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Abstract

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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Acknowledgements

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.

Funding

This work was also supported by FAPESP #2012/23645-4 (Dr. Paulo Cesar Mayorca) and FAPESP #2013/21719–3 (Dr. Maurício Lacerda Nogueira).

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Authors and Affiliations

  1. Vector-Borne Diseases Laboratory, Virology Center, Adolfo Lutz Institute, São Paulo, Brazil

    Mariana Sequetin Cunha & Giovana Santos Caleiro

  2. Enteric Diseases Laboratory, Virology Center, Adolfo Lutz Institute, São Paulo, Brazil

    Adriana Luchs

  3. Respiratory Diseases Laboratory, Virology Center, Adolfo Lutz Institute, São Paulo, Brazil

    Fabiana Cristina Pereira dos Santos

  4. Virology Laboratory, São Paulo Institute of Tropical Medicine, University of São Paulo, São Paulo, Brazil

    Giovana Santos Caleiro

  5. Faculdade de Medicina de São Jose do Rio Preto (FAMERP), São Paulo, SP, Brazil

    Maurício Lacerda Nogueira

  6. Department of Pathology, School of Veterinary Medicine and Animal Sciences, University of São Paulo, São Paulo, Brazil

    Paulo César Maiorka

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  1. Mariana Sequetin Cunha
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Contributions

M.S.C. and P.C.M. conceived the study and designed the study protocol; M.S.C., A.L., F.C.P.S., J.S.N., A.Y.M, M.L.N., and P.C.M. participated in conducting the study; M.S.C., J.N.S. and A.Y.M. acquired the data. M.S.C. performed the plaque, TCID50 and IFA assays; M.S.C., G.S.C. and F.C.P.S. performed the RT-qPCR assays; M.S.C. and A.L. conducted the sequencing assays; M.S.C., A.L., F.C.P.S., M.L.N. and P.C.M. analyzed and interpreted the data; M.S.C. and A.L. drafted the manuscript; F.C.P.S, J.S.N., G.S.C., A.Y.M., M.L.N. and P.C.M. critically revised the manuscript for intellectual content. All authors read and approved the final version. M.S.C. is the guarantor of the paper.

Corresponding author

Correspondence to Mariana Sequetin Cunha.

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Ethical approval

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

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  • DOI: https://doi.org/10.1007/s00705-020-04680-w

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