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Updated: Aug 20, 2025
In Vitro Selection of Aptamers to Differentiate Infectious from Non-Infectious Viruses
Published on: September 7, 2022
Foot-and-mouth disease virus: DNA aptamer selection for the 3ABC protein
Camila Maria de Sousa Lacerda1, Nathalie Bonatti Franco Almeida1, Viviane Cristina Fernandes Dos Santos1
1Centro de Desenvolvimento da Tecnologia Nuclear, CDTN, Belo Horizonte, MG 31270-901, Brasil; MedicOnChip, Parque Tecnológico de Belo Horizonte-BH-TEC, Belo Horizonte, MG, Brasil.
Abstract:
Foot-and-mouth disease (FMD) is a devastating livestock disease caused by foot-and-mouth disease virus (FMDV), a member of the Picornaviridae family. The 3ABC is a non-structural protein of FMDV, produced during viral replication and absent from inactivated FMD vaccines. Nucleic acid aptamers are DNA or RNA oligonucleotides capable of binding with high specificity and affinity to a molecular target. The aim of this study was to obtain DNA aptamers specific for 3ABC protein with a view of their application in the FMD diagnosis. Aptamers are usually obtained through SELEX (Systematic Evolution of Ligands by EXponential enrichment) procedure. In this study, an aptamer (termed FMDV1) was selected by a variation of this technique called Capillary Electrophoresis SELEX (CE-SELEX). The FMDV1 aptamer showed high binding affinity to the 3ABC protein with Kd value in the nano molar range: 22.69 ± 1.79 nM. The FMDV1 aptamer binding to 3ABC was significantly higher when compared with the BSA protein, used as control, demonstrating its specificity.
Insights
Researchers developed a DNA aptamer, FMDV1, to detect the foot-and-mouth disease virus (FMDV) 3ABC protein. This specific aptamer shows high binding affinity, paving the way for improved FMD diagnostics.
Area of Science:
- Veterinary Virology
- Molecular Biology
- Biotechnology
Background:
- Foot-and-mouth disease (FMD) is a highly contagious livestock disease caused by the foot-and-mouth disease virus (FMDV).
- The FMDV 3ABC protein is crucial for viral replication and is absent in inactivated vaccines, making it a potential diagnostic target.
- Nucleic acid aptamers offer high specificity and affinity for molecular targets, presenting an alternative to traditional diagnostic methods.
Purpose of the Study:
- To isolate and characterize DNA aptamers that specifically bind to the FMDV 3ABC non-structural protein.
- To evaluate the potential application of these aptamers in developing novel diagnostic tools for FMD.
Main Methods:
- Selection of DNA aptamers using a modified Systematic Evolution of Ligands by EXponential enrichment (SELEX) technique, specifically Capillary Electrophoresis SELEX (CE-SELEX).
- Characterization of aptamer binding affinity and specificity to the 3ABC protein using quantitative binding assays.
- Utilized Bovine Serum Albumin (BSA) as a control protein to assess specificity.
Main Results:
- A specific DNA aptamer, designated FMDV1, was successfully selected for the FMDV 3ABC protein.
- The FMDV1 aptamer demonstrated high binding affinity to the 3ABC protein, with a dissociation constant (Kd) in the nanomolar range (22.69 ± 1.79 nM).
- The binding of FMDV1 to 3ABC was significantly greater than to BSA, confirming its high specificity.
Conclusions:
- The FMDV1 DNA aptamer is a highly specific and high-affinity binder for the FMDV 3ABC protein.
- This aptamer holds promise for the development of sensitive and specific diagnostic assays for foot-and-mouth disease.
- Aptamer-based diagnostics could offer advantages in FMD detection and surveillance.
