Selection and Characterization of Single-Stranded DNA Aptamers of Diagnostic Potential against the Whole Zika Virus

Abstract

Zika virus became a major public health problem in early 2015, when cases of Guillain–Barré syndrome and microcephaly were associated with viral infection. Currently, ZIKV is endemic in all tropical areas of the world, and the chance for future Zika epidemics remains very real and accurate diagnosis is crucial. The aim of this work was to select specific ssDNA aptamers that bind to the entire Zika virus and can be used to compose specific diagnostics, without cross-reactivity with other flaviviruses. Zika virus was cultivated in Vero cells and used as a target for aptamer selection. Aptamers specific for the ZIKV were selected using whole-virus SELEX, with counterselection for other flavivirus. Secondary and tertiary structures were evaluated and the molecular anchoring between the aptamers and target were simulated by the HDOCK server. Aptamer interaction was evaluated by ELISA/ELASA and the dissociation constant (Kd) was calculated by thermophoresis. Four ZIKV-specific aptamers were selected. The best two were further characterized and proved to be specific for ZIKV. Aptamers are capable of binding specifically to the ZIKV and differentiate from Dengue virus. The aptamers selected in this work can be used as capture agents in the composition of diagnostic tests to specifically detect ZIKV infection.

Moving towards Personalized Medicine—The Broad Use of Aptamers for Targeted Theranostic

“…They can be used in combination with high-throughput sequencing technologies, such as 16S rRNA sequencing to identify and quantify specific microbial populations in the gut microbiota and to identify and quantify specific microbial metabolites, which are important for gut health [118]. This approach allows researchers to better understand the interactions between the microbiome and the host and how they influence overall health and disease, providing a more accurate diagnosis and better understanding of the disease [119][120][121][122][123].…”

“…For example, Wandtke et al (2015) designed aptamers to selectively bind to viral proteins or other molecules involved in viral replication and infection with high specificity and sensitivity, preventing their replication or disrupting their functions [124]. Here, the authors discuss several examples of aptamer-based assays developed for the detection of hepatitis C and Human Immunodeficiency Virus (HIV) and also how this novel method could be used as a therapeutic agent for viral infections, such as influenza, with A-20 and V46 aptamers [119,120], dengue fever, with S15G3 aptamer [121], and Zika virus [122]. Additionally, aptamers can be used to deliver therapeutic molecules, such as antibiotics, antivirals, antifungals, or probiotics, specifically to the site of infection or dysbiosis in the human microbiota, reducing side effects and increasing efficacy [123].…”