核酸适配体联合流式细胞术定量检测创伤弧菌方法的建立
Methodology establishment of detection of Vibrio vulnificus by flow cytometry and aptamer
刘德婧 1胡波 1彭定发 1欧阳生群 1王梁华 1焦炳华1
作者信息
- 1. 中国人民解放军第二军医大学基础部 生物化学与分子生物学教研室,上海 200433
- 折叠
摘要
目的 建立快速、准确定量检测创伤弧菌的方法.方法 将修饰有5''-FITC荧光标签的,能够与创伤弧菌高亲和力,强特异性结合的核酸适配体与流式细胞技术联合,实现创伤弧菌的快速检测.通过对不同浓度创伤弧菌菌液的重复检测,测定该检验方法的空白限、检测限、线性范围等.结果 核酸适配体联合流式细胞技术可快速检测创伤弧菌,检测时长＜1 h,检测下限最低可达29 CFU/mL.结论 创伤弧菌适配体是优秀的检测元件,流式细胞技术可以实现准确的定量检测,该检测方法有很大的应用潜力.
Abstract
Objective Establish detection method to measure Vibrio vulnificus rapidly and accurately.Methods Using flow cytometry(FCM)and a 5'-FITC fluorescent labeled aptamer with high binding affinity to detect Vibrio vulnificus rapidly.Measure a series of concentrations of Vibrio vulnificus to identify the Limit of Blank, Lower Limit of Detection, Linearity Range, etc.Results Combined application of FCM and the aptamer can detect Vibrio vulnificus rapidly with the duration less than 1 hour and lower limit of detection as low as 29 CFU/mL.Conclusion The aptamer targeting Vibrio vulnificus is an excellent detective element, while FCM can realize accurate quantitative detection.The detection method has great application potential.
关键词
核酸适配体/创伤弧菌/流式细胞技术/病原微生物检测Key words
aptamer/Vibrio vulnificus/flow cytometery/pathogen detection引用本文复制引用
基金项目
国家高技术研究发展计划(2013AA092904)
出版年
2017
NETL
NSTL
维普
万方数据