Guangxi Key Laboratory of Marine Natural Products and Combinatorial Biosynthesis Chemistry, Guangxi Academy of Sciences, Nanning, China.
College of Life Science, Henan Normal University, Xinxiang, China.
J Fish Dis. 2019 Nov;42(11):1523-1529. doi: 10.1111/jfd.13066. Epub 2019 Aug 26.
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 × 10 /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.
作为海洋动物和人类的主要机会致病菌，Alginate-degrading Vibrio (Vibrio alginolyticus) 给水产养殖业造成了巨大的经济损失。在我们之前的研究中，通过指数富集配体系统进化技术 (SELEX) 生成了针对活 V. alginolyticus 的 ssDNA 适体 VA2。在本研究中，我们首次开发了基于适体 (VA2) 的酶联适体吸附测定法 (VA2-ELASA)，用于快速检测水产养殖病原体 V. alginolyticus。VA2-ELASA 可以实现对 V. alginolyticus 感染的快速检测，具有高度特异性和敏感性。VA2-ELASA 可以特异性识别 V. alginolyticus，但不能识别其他非靶标细菌株。VA2-ELASA 可以检测到浓度为 5×10/ml 的 V. alginolyticus，孵育时间短至 1 分钟，孵育温度高达 45°C，这证明了本研究中新型 VA2-ELASA 的灵敏度和稳定性。VA2-ELASA 完成检测过程不到一个小时。特异性、灵敏度和易操作性的特点使 VA2-ELASA 成为水产养殖中快速诊断病原体 V. alginolyticus 的一种新的有用技术。
