1. Development and validation of RT-LAMP for detecting yellow fever virus in non-human primates samples from Brazil.
- Author
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Cardoso SF, Yoshikawa AAG, Pinheiro IC, Granella LW, Couto-Lima D, Neves MSAS, Mansur DS, Pitaluga AN, and Rona LDP
- Subjects
- Animals, Brazil epidemiology, Molecular Diagnostic Techniques methods, Primates virology, Yellow fever virus genetics, Yellow fever virus isolation & purification, Yellow Fever diagnosis, Yellow Fever virology, Yellow Fever epidemiology, Sensitivity and Specificity, Nucleic Acid Amplification Techniques methods
- Abstract
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).)
- Published
- 2024
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