Fast and visual RT-LAMP assay for detection of oropouche virus

Abstract

Background: The epidemic potential of Oropouche virus (OROV) highlights the urgent need for rapid and accessible diagnostic methods to improve testing capacity and reduce result turnaround times, especially in tropical regions where OROV prevalence is rising. Objective: To develop and validate a rapid, cost-effective diagnostic assay for OROV detection suitable for use in resource-limited and field settings. Methods: A reverse transcription loop-mediated isothermal amplification (RT-LAMP) assay was designed and optimized for OROV detection. The assay's performance was evaluated in terms of sensitivity, specificity, time to result, and ease of visual interpretation based on colorimetric changes. Results: The RT-LAMP assay produced results within 30 to 35 minutes and enabled straightforward visual detection through a clear color change. Its detection limit was comparable to that of real-time PCR, the current gold standard for molecular diagnosis. The assay's simplicity and minimal equipment requirements make it well-suited for decentralized testing environments. Conclusions: This RT-LAMP assay represents a rapid, sensitive, and accessible tool for OROV diagnosis, with significant potential to enhance outbreak response and surveillance efforts in endemic regions.