Introduce a novel, extremely sensitive aptamer against staphylococcal enterotoxin type D

Abstract

Background: Staphylococcus aureus (S. aureus) is a globally prevalent foodborne pathogen responsible for significant public health concerns. Staphylococcal food poisoning (SFP) results from staphylococcal enterotoxins (SEs) produced by specific strains of S. aureus. Rapid and effective detection of SEs remains a significant challenge for public health authorities. Aptamers, short single-stranded DNA(ssDNA), RNA, or synthetic xeno nucleic acid (XNA) molecules, exhibit high affinity for binding to their specific targets. Due to their unique properties, including low production costs, ease of chemical modification, high thermal stability, and reproducibility, aptamers present a viable alternative to antibodies for diagnostic and therapeutic applications.

Objectives: This research aimed to isolate high-affinity ssDNA aptamers with specificity for staphylococcal enterotoxin D (SED).

Methods: The systematic evolution of ligands by the exponential enrichment (SELEX) method was utilized to identify specific aptamers. These aptamers were then validated using enzyme-linked apta-sorbent assay (ELASA) and surface plasmon resonance (SPR) to assess their binding characteristics and affinity.

Results: SELEX successfully identified aptamers with strong binding affinity to SED. Among the identified candidates, one aptamer, Aptamer 1, exhibited the highest specificity for SED with a dissociation constant (K_D) of 4.4 ± 2.26 nM. The limit of detection (LOD) for SED using this aptamer was determined to be 45 nM.

Conclusions: The findings indicate that the ELASA system designed using the aptamer developed in this study demonstrates higher specificity, sensitivity, and reproducibility in detecting enterotoxin D. This novel aptamer offers significant potential for applications in diagnostic platforms targeting S. aureus enterotoxins.

Keywords: ELASA; SELEX; SPR; Staphylococcal enterotoxin D; Staphylococcus aureus; ssDNA-aptamer.