Development and validation of RT-LAMP for detecting yellow fever virus in non-human primates samples from Brazil.

Autor: Cardoso SF; Department of Cell Biology, Embryology, and Genetics, Federal University of Santa Catarina (UFSC), Florianópolis, Brazil.; Directorate of Epidemiological Surveillance (DIVE), Santa Catarina's State Health Secretary, Florianópolis, Brazil., Yoshikawa AAG; Department of Cell Biology, Embryology, and Genetics, Federal University of Santa Catarina (UFSC), Florianópolis, Brazil., Pinheiro IC; Department of Cell Biology, Embryology, and Genetics, Federal University of Santa Catarina (UFSC), Florianópolis, Brazil., Granella LW; Department of Microbiology, Immunology and Parasitology, Federal University of Santa Catarina (UFSC), Florianópolis, Brazil., Couto-Lima D; Hematozoan Transmitting Mosquito Laboratory, Oswaldo Cruz Institute (IOC), FIOCRUZ, Rio de Janeiro, RJ, Brazil., Neves MSAS; Hematozoan Transmitting Mosquito Laboratory, Oswaldo Cruz Institute (IOC), FIOCRUZ, Rio de Janeiro, RJ, Brazil., Mansur DS; Department of Microbiology, Immunology and Parasitology, Federal University of Santa Catarina (UFSC), Florianópolis, Brazil., Pitaluga AN; Oswaldo Cruz Institute (IOC), FIOCRUZ, Rio de Janeiro, RJ, Brazil. pitaluga@ioc.fiocruz.br.; National Institute of Science and Technology in Molecular Entomology, National Council for Scientific and Technological Development (INCT-EM, CNPq), Rio de Janeiro, Brazil. pitaluga@ioc.fiocruz.br., Rona LDP; Department of Cell Biology, Embryology, and Genetics, Federal University of Santa Catarina (UFSC), Florianópolis, Brazil. luisa.rona@ufsc.br.; National Institute of Science and Technology in Molecular Entomology, National Council for Scientific and Technological Development (INCT-EM, CNPq), Rio de Janeiro, Brazil. luisa.rona@ufsc.br.
Jazyk: angličtina
Zdroj: Scientific reports [Sci Rep] 2024 Sep 28; Vol. 14 (1), pp. 22520. Date of Electronic Publication: 2024 Sep 28.
DOI: 10.1038/s41598-024-74020-4
Abstrakt: Monitoring yellow fever in non-human primates (NHPs) is an early warning system for sylvatic yellow fever outbreaks, aiding in preventing human cases. However, current diagnostic tests for this disease, primarily relying on RT-qPCR, are complex and costly. Therefore, there is a critical need for simpler and more cost-effective methods to detect yellow fever virus (YFV) infection in NHPs, enabling early identification of viral circulation. In this study, an RT-LAMP assay for detecting YFV in NHP samples was developed and validated. Two sets of RT-LAMP primers targeting the YFV NS5 and E genes were designed and tested together with a third primer set to the NS1 locus using NHP tissue samples from Southern Brazil. The results were visualized by colorimetry and compared to the RT-qPCR test. Standardization and validation of the RT-LAMP assay demonstrated 100% sensitivity and specificity compared to RT-qPCR, with a detection limit of 12 PFU/mL. Additionally, the cross-reactivity test with other flaviviruses confirmed a specificity of 100%. Our newly developed RT-LAMP diagnostic test for YFV in NHP samples will significantly contribute to yellow fever monitoring efforts, providing a simpler and more accessible method for viral early detection. This advancement holds promise for enhancing surveillance and ultimately preventing the spread of yellow fever.
(© 2024. The Author(s).)
Databáze: MEDLINE
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