Development of novel aptamer‐based enzyme‐linked apta‐sorbent assay (ELASA) for rapid detection of mariculture pathogen Vibrio alginolyticus

Abstract

As the major opportunistic pathogen to both marine animals and humans, Vibrio alginolyticus (V. alginolyticus) has caused heavy economic losses to mariculture. ssDNA aptamer VA2 targeting live V. alginolyticus was generated by systematic evolution of ligands by exponential enrichment (SELEX) technology in our previous study. In this study, we first developed aptamer (VA2)‐based enzyme‐linked apta‐sorbent assay (VA2‐ELASA) for rapid detection of mariculture pathogen V. alginolyticus. The VA2‐ELASA could achieve the rapid detection for V. alginolyticus infection with high specificity and sensitivity. The VA2‐ELASA could specifically identify V. alginolyticus, but not other non‐target bacterial strains. VA2‐ELASA could detect V. alginolyticus at the concentration of 5 × 104/ml, the incubation time short to 1 min and the incubation temperature as high as 45°C, which proved sensitivity and stability of the novel VA2‐ELASA in this study. It took less than one hour to accomplish the detection process by VA2‐ELASA. The characteristics of specificity, sensitivity and easy operation make VA2‐ELASA a novel useful technology for the rapid diagnosis of pathogen V. alginolyticus in mariculture.

Application of MALDI-TOF MS Profiling Coupled With Functionalized Magnetic Enrichment for Rapid Identification of Pathogens in a Patient With Open Fracture

“…V. alginolyticus, a type of Gram-negative opportunistic bacteria, infects both humans and aquatic animals (Xie et al, 2020). This infection is one of the main causes of aquaculture disease resulting in economic losses in mariculture in South China (Xie et al, 2005;Yu et al, 2018;Yu et al, 2019). Therefore, it was not surprising that V. alginolyticus was identified in this case.…”

A Novel Method for Sensitive Detection of Vibrio alginolyticus Based on Aptamer and Hybridization Chain Reaction in Aquaculture

“…Adaptor valence refers to the number of ligands that can attach a receptor to an entity. Previous reports have highlighted that increasing the valence state of ligands on a multivalent scaffold significantly enhances the binding affinity of the multi-Apt [18]. Based on our previous investigation, the optimal binding avidity of multi-Apt was observed when the neighboring aptamer distance was set at 72 bp, with a trigger hairpin ratio of 50:1 [26].…”

“…The developed ELISA demonstrated the ability to detect V. alginolyticus at a concentration as low as 5 × 10 4 CFU/mL. An important achievement was the substantial reduction in the incubation time to just 1 min while maintaining accurate results even at a high incubation temperature of 45 • C [18]. A differential fluorescence method for Vibrio anguillarum detection in seawater and tissue samples was developed based on aptamer, and the limit of detection (LOD) of V. anguillarum reached 10 2 CFU/mL, with a good linear relationship was found in the detection range of 10 2 ~10 8 CFU/mL [19].…”