Yellow Fever Virus (YFV), a mosquito-borne flavivirus, continues to pose a significant public health threat in tropical and subtropical regions, particularly in Africa and South America (Malik et al., 2023). Despite the availability of an effective vaccine, outbreaks persist due to factors such as low vaccination coverage, urbanization, climate change, and vaccine hesitancy.

YFV is part of the Flaviviridae family, which includes other medically important pathogens like dengue virus (DENV), Zika virus (ZIKV), and West Nile virus (WNV). These viruses share antigenic similarities, leading to cross-reactivity in serological assays and complicating accurate differential diagnosis. Additionally, the current gold-standard diagnostic methods—such as ELISA, RT-PCR, and virus isolation—require specialized infrastructure and trained personnel, which limits their accessibility in resource-limited settings (Madere et al., 2025). This diagnostic challenge is particularly acute in regions where multiple flaviviruses co-circulate, and where access to advanced diagnostic tools is limited.

In response to these challenges, there is a growing interest in the development of molecular tools that are specific, stable, and adaptable to point-of-care use. Aptamers, single-stranded DNA or RNA oligonucleotides selected through the Systematic Evolution of Ligands by Exponential Enrichment (SELEX) process (Ellington and Szostak, 1990, Tuerk and Gold, 1990), have emerged as promising alternatives or complements to antibodies in diagnostic applications. Aptamers exhibit high affinity and specificity for their targets, are synthetically produced, thermally stable, and can be modified to enhance their functionality (Kong and Byun, 2013). These properties make aptamers particularly attractive for the development of low-cost, portable diagnostic platforms suitable for application in endemic regions.

Given the continued burden of YFV and other flaviviruses, especially in underserved areas, there is a critical need to advance innovative diagnostic solutions. In recent years, aptamers have been successfully developed against several viral targets, including flaviviral antigens, demonstrating their utility in biosensors, lateral flow assays, and other diagnostic formats (Argondizzo et al., 2020, Blázquez and Jiménez de Oya, 2024).

Nonstructural glycoprotein 1 (NS1) is a key antigen of flaviviruses. It is found in both membrane-associated and soluble secreted forms. NS1 plays a vital role in viral replication and significantly contributes to the modulation of the host immune response (Fisher et at., 2023). Due to its diagnostic importance, NS1 was selected as the target for the development of novel DNA aptamers specific to the YFV.

This study investigates the use of aptamer-based strategies to improve the specificity and effectiveness of diagnostic tools for Yellow Fever and other related flaviviruses, aiming to address a critical need in global health diagnostics.