特异性检测卵形鲳鲹源哈维氏弧菌的核酸适配体的筛选与鉴定

哈维氏弧菌(Vibrio harveyi)是海水养殖动物常见的主要致病菌之一,严重危害着我国华南沿海地区水产养殖业的健康可持续发展。针对哈维氏弧菌的快速检测诊断技术和抑菌技术等方面进行系统研究,并开发具有市场发展潜力的快速检测试剂盒和抗菌功能产品,对控制哈维氏弧菌的危害极其重要。为此,本研究以哈维氏弧菌为靶标,利用指数富集的配基系统进化(Systematic Evolution of Ligands by Exponential Enrichment, SELEX)技术,筛选特异性识别哈维氏弧菌的核酸适配体,并对核酸适配体的性质进行系统性研究。结果显示,核酸适配体H10、H13能高特异性和高亲和性地识别哈维氏弧菌,并且无毒副作用。同时,靶标性质分析表明核酸适配体(H10,H13)的靶标可能是细胞膜蛋白或膜蛋白相关成分。本研究运用SELEX技术筛选获得的ssDNA核酸适配体H10和H13,具有高特异性和亲和性,可用于研发操作便捷、灵敏度高的哈维氏弧菌快速检测技术。

Selection and Identification of Aptamers Specifically Recognizing Pathogenic Vibrio harveyi Isolated from Cultured Golden Pompano (Trachinotus ovatus)

Vibrio harveyi is one of the main pathogenic bacteria that are common in marine aquaculture animals, which seriously endangers the healthy and sustainable development of the aquaculture industry in the coastal areas of South China. Systematic research on the rapid detection and diagnosis technology and antibacterial technology against V. harveyi, and the development of rapid detection kits and antibacterial products with market development potential are extremely important to control the harm of V. harveyi. This study took V. harveyi as the target and used the Systematic Evolution of Ligands by Exponential Enrichment (SELEX) technology to screen for aptamers that specifically recognize V. harveyi, and systematically study the properties of aptamers. The results showed that the selected aptamers H10 and H13 could recognize target V. harveyi with high specificity and affinity without cytotoxicity. The target properties analysis results showed that the targets of aptamers H10 and H13 might be cell membrane proteins or membrane proteins related components. The ssDNA aptamers H10 and H13 selected by SELEX technology have high specificity and affinity targeting V. harveyi, which could be used to develop the rapid detection technology against V. harveyi with convenient operation and high sensitivity.