基于适配体的金黄色葡萄球菌流式细胞术检测方法  

Detection of Staphylococcus aureus with Flow Cytometry and Aptamer

作　　者：董晓琳[1] 李志萍[1] 高玮村 樊洪超 王园园[1] 吴永魁[1] 夏志平[1] 

机构地区：[1]军事医学科学院军事兽医研究所/吉林省人兽共患病预防与控制重点实验室,长春130122

出　　处：《东北农业科学》2016年第3期81-86,共6页Journal of Northeast Agricultural Sciences

基　　金：武汉东湖新技术开发区3551光谷人才计划-第七批

摘　　要：本研究以金黄色葡萄球菌为对象,旨在建立一种基于荧光标记核酸适配体的流式细胞术检测方法。通过优化流式细胞仪检测参数、适配体用量及折叠方式,观察适配体针对不同方式处理细菌的识别效果建立检测方法,并对该方法的特异性和灵敏性进行了验证。结果显示:不经高温变性处理的适配体检测结果最佳,适配体最佳使用浓度为250 nmol/L,对活菌识别效果最强。本实验建立的基于适配体的流式细胞术检测方法特异性强、过程简单,40 min于混合细菌中特异性检测、鉴别靶细菌,为金黄色葡萄球菌的检测提供了一种有效方法。The aim of this study is to establish a flow cytometry method based on fluorescent-labeled nucleic acid aptamer for detection of Staphylococcus aureus. By optimizing flow cytometry parameters, the amount and folding methods of the aptamer and bacterial treatment, specificity and sensitivity of this method was finally verified. The resuits showed that aptamer which was not folding by the high temperature denaturation process has a good identifica- tion and the strongest binding with viable bacteria, the optimal working concentrations of aptamer is 250 nmol/L. We determined Staphylococcus aureus after pre-incubation in a total analysis time of 40 min, even in the presence of Escherichia coli cells. The results proved that flow cytometry combined with fluorescently labeled aptamer offers advantages of speed and specificity for the detection of Staphylococcus aureus.

关 键 词：适配体 流式细胞术 金黄色葡萄球菌 

分 类 号：S851.3[农业科学—预防兽医学]