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朱冬琳,余庆,刘明珠,肖贺贺,吴思婷,牙韩争,覃仙玲,陈宪云,董德信,聂振平,肖俊,王一兵,李鹏飞.基于核酸适配体吸附检测技术的卵形鲳鲹源溶藻弧菌的快速检测[J].广西科学院学报,2019,35(3):219-224. [点击复制]
- ZHU Donglin,YU Qing,LIU Mingzhu,XIAO Hehe,WU Siting,YA Hanzheng,QIN Xianling,CHEN Xianyun,DONG Dexin,NIE Zhenping,XIAO Jun,WANG Yibing,LI Pengfei.Establishment and Characterization of Enzyme-Linked Apta-Sorbent Assay (ELASA) for Rapid Diagnosis of Vibrio alginolyticus Infection in Cultured Trachinotus ovatus[J].Journal of Guangxi Academy of Sciences,2019,35(3):219-224. [点击复制]
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| 基于核酸适配体吸附检测技术的卵形鲳鲹源溶藻弧菌的快速检测 |
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朱冬琳1, 余庆1, 刘明珠2, 肖贺贺2,3, 吴思婷2, 牙韩争1, 覃仙玲1, 陈宪云1, 董德信1, 聂振平2, 肖俊4, 王一兵5, 李鹏飞2,6
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| (1.广西科学院, 广西近海海洋环境科学重点实验室, 广西南宁 530007;2.广西壮族自治区海洋研究所, 广西海洋生物技术重点实验室, 广西北海 536000;3.河南师范大学生命科学学院, 河南新乡 453007;4.广西壮族自治区水产科学研究院, 广西水产遗传育种与健康养殖重点实验室, 广西南宁 530021;5.广西民族大学海洋与生物技术学院, 广西南宁 530006;6.广西海洋天然产物与组合生物合成化学重点实验室, 广西南宁 530007) |
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| 摘要: |
| 溶藻弧菌是引起广西等华南沿海地区海水养殖鱼类发生细菌性鱼病的主要致病菌之一，其引起的鱼病具有发病迅速、死亡率高、流行面广等特点，严重威胁着华南地区水产养殖业的健康可持续发展。着力发展操作便捷、成本低、耗时短、准确度高的水产致病菌快速检测技术，对于及早发现、确定病原，进而有的放矢地制定治疗方案来控制病原扩散、降低损失意义重大。在先前研究中，我们基于指数富集的配基系统进化技术（Systematic evolution of ligands by exponential enrichment technology，SELEX）筛选获得特异性识别卵形鲳鲹源溶藻弧菌的核酸适配体，本研究中我们基于核酸适配体VA8开展溶藻弧菌的快速检测诊断技术的研究，开发出一种新型的能够快速检测溶藻弧菌的核酸适配体吸附检测技术（Aptamer VA8-based enzyme-linked apta-sorbent assay，VA8-ELASA），并对VA8-ELASA技术检测溶藻弧菌的特异性和灵敏性进行分析研究。VA8-ELASA技术可以用于溶藻弧菌的快速检测，具有特异性强、灵敏度高的特点。本研究基于核酸适配体VA8建立的新型核酸适配体吸附技术（VA8-ELASA），有望实现对广西卵形鲳鲹养殖中溶藻弧菌病的快速诊断、实时监控和有效预防。 |
| 关键词: 溶藻弧菌 卵形鲳鲹 ELASA SELEX VA8 特异性 灵敏性 快速诊断 |
| DOI：10.13657/j.cnki.gxkxyxb.20190903.010 |
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| 基金项目:广西自然科学基金项目（2017GXNSFBA198176，2018JJB150002），广西重点研发计划（2018AB52003），广西科技人才专项（2018AD19367），现代农业产业技术体系专项资金项目（CARS-46）和广西科学院基本科研业务费项目（2018YJJ903，2018YJJ902）资助。 |
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| Establishment and Characterization of Enzyme-Linked Apta-Sorbent Assay (ELASA) for Rapid Diagnosis of Vibrio alginolyticus Infection in Cultured Trachinotus ovatus |
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ZHU Donglin1, YU Qing1, LIU Mingzhu2, XIAO Hehe2,3, WU Siting2, YA Hanzheng1, QIN Xianling1, CHEN Xianyun1, DONG Dexin1, NIE Zhenping2, XIAO Jun4, WANG Yibing5, LI Pengfei2,6
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| (1.Guangxi Key Laboratory of Marine Environmental Science, Guangxi Academy of Sciences, Nanning, Guangxi, 530007, China;2.Guangxi Key Laboratory for Marine Biotechnology, Guangxi Institute of Oceanography, Beihai, Guangxi, 536000, China;3.College of Life Science, Henan Normal University, Xinxiang, Henan, 453007, China;4.Guangxi Key Laboratory of Aquatic Genetic Breeding and Healthy Breeding, Guangxi Academy of Fishery Science, Nanning, Guangxi, 530021, China;5.School of Marine Sciences and Biotechnology, Guangxi University for Nationalities, Nanning, Guangxi, 530006, China;6.Guangxi Key Laboratory of Marine Natural Products and Combinatorial Biosynthesis Chemistry, Nanning, Guangxi, 530007, China) |
| Abstract: |
| Vibrio alginolyticus is one of the main pathogens causing bacterial fish disease in marine fish in coastal area of South China including Guangxi,and its fish disease has the characteristics of rapid onset,high mortality and wide popularity,which seriously threatens the healthy and sustainable development of aquaculture in South China.Focusing on developing rapid detection techniques for aquatic pathogens with convenient operation,low cost,short time and high accuracy are of great significance for early detection and identification of pathogens and then targeting to develop a treatment plan to control the spread of pathogens and reduce losses.In previous studies,ssDNA aptamers against V.alginolyticus were screened based on systematic evolution of ligands by exponential enrichment technology (SELEX).In this study,we carried out a research on a rapid detection and diagnosis technology of V.alginolyticus based on aptamer VA8,and developed a new type of Aptamer VA8-based enzyme-linked aptasorbent assay (VA8-ELASA) for rapid detection of V.alginolyticus.The specificity and sensitivity of VA8-ELASA technology for the detection of V.alginolyticus were analyzed.VA8-ELASA technology can be used for the rapid detection of V.alginolyticus with strong specificity and high sensitivity.The novel VA8-ELASA technology based on aptamer VA8 is expected to realize rapid diagnosis,real-time monitoring and effective prevention of V.alginolyticus disease in Trachinotus ovatus aquaculture in Guangxi. |
| Key words: V.alginolyticus Trachinotus ovatus ELASA SELEX VA8 specificity sensitivity rapid diagnosis |
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