利用糖芯片技术快速检测沙门氏菌

Rapid Detection of Salmonella Using Carbohydrate Microarrays

为了探讨糖芯片在沙门氏菌检测上的应用,以沙门氏菌ATCC31685为主要研究对象,制作不同类型的糖芯片用于优化糖芯片检测条件;在优化条件的基础上,降低糖浓度或细菌浓度用于确定糖芯片检测限值;继而系统地研究了糖芯片在沙门氏菌和凝集素FimH拮抗剂筛选上的应用,并探究糖芯片对菌株的破坏性。由实验现象可知,沙门氏菌可与甘露糖化合物修饰的糖芯片结合,其中,以Man-1与NHS玻片所构建的糖芯片效果最优,并且两种不同的沙门氏菌(ATCC31685和ATCC9184)与糖芯片的结合能力不同,因此糖芯片可作为沙门氏菌的一种快速检测手段;在糖浓度为313 mmol/L或细菌浓度106个/mL时荧光信号接近背景值,并且糖芯片检测后对菌株无破坏性,可继续其他检测项目;此外,实验结果表明糖芯片可作为FimH蛋白拮抗剂的筛选工具。

The applicability of carbohydrate microarrays for Salmonella detection was investigated, using Salmonella ATCC31685 as the primary test subject. Different types of carbohydrate microarrays were first tested to optimize the detection conditions. Then, using the optimized conditions, decreased carbohydrate concentrations or bacterial concentrations were used to determine the microarray＇s limit of detection. The carbohydrate microarrays were tested for their destructiveness of bacterial strains as well as their ability to screen for antagonists of Salmonella lectin FimH. The results indicated that Salmonella bound to modified mannose compounds in the carbohydrate microarrays, among which the microarray chips with Man-1 and NHS were the most optimal In addition, the microarray-binding abilities of two Salmonella strains （ATCC31685 and ATCC9184） were found to be different. Therefore, carbohydrate microarrays can serve as a rapid procedure for detecting Salmonella. At a carbohydrate concentration of 313 mmol/L or a cell concentration of 106/mL, the fluorescent signal approaches a background value. The carbohydrate microarray detection of Salmonella is non-destructive and can be used continuously. In addition, these microarrays can be used as tools for screening FimH protein antagonists.