State Key Laboratory of Pulp and Paper Engineering, School of Light Industry and Food, South China University of Technology, Guangzhou 510640, PR China.
J Biomater Sci Polym Ed. 2011;22(14):1881-93. doi: 10.1163/092050610X528570. Epub 2010 Oct 19.
A fast and sensitive method based on fluorescent carboxymethyl chitosan/CdS quantum dots (CMCS-CdS QDs) composites was developed for specific detection of Staphylococcus aureus in food and the environment. Fluorescent CMCS-CdS QDs were prepared in aqueous solution through a green method. A human immunoglobulin (IgG) antibody was then bioconjugated to the QDs in the presence of 1-ethyl-3-(3)-dimethylaminopropyl carbodiimide hydrochloride (EDC) and N-hydroxysuccinimide (NHS) to make a novel type of mono-dispersed water-soluble fluorescent bioprobes. The fluorescent bioprobes were employed to identify S. aureus by incubating them with the bacteria for a certain time and observing the marked cells under fluorescence microscopy after removing free fluorescent QDs. Fluorescence microscopy images showed the S. aureus cells were successfully recognized by the bioprobes. Several other bacteria commonly found in environment such as Escherichia coli and Bacillus subtilis were also incubated with the bioprobes to test their specificity. It was found that the novel QDs-CMCS-IgG bioprobes had specific identification to S. aureus cells. Fluorescence measurement using a luminescence spectrometer could be applied to quantify S. aureus cells. The fluorescence intensity of the samples at 600 nm was proportional to the cell concentration in the range of 10(3)-10(7) cfu/ml, and the detection limit was as low as 900 cfu/ml. Considering the simplicity and cost-efficiency of this method, its application in the identification and quantification of bacteria in clinical, food and environmental samples is anticipated.
一种基于荧光羧甲基壳聚糖/CdS 量子点(CMCS-CdS QDs)复合材料的快速灵敏方法被开发出来,用于特异性检测食品和环境中的金黄色葡萄球菌。通过绿色方法在水溶液中制备荧光 CMCS-CdS QDs,然后在 1-乙基-3-(3)-二甲基氨基丙基碳化二亚胺盐酸盐(EDC)和 N-羟基琥珀酰亚胺(NHS)的存在下将人免疫球蛋白(IgG)抗体与 QDs 生物偶联,制得新型单分散水溶性荧光生物探针。通过将荧光生物探针与细菌孵育一段时间,并用荧光显微镜观察去除游离荧光 QDs 后的标记细胞,来识别金黄色葡萄球菌。荧光显微镜图像显示生物探针成功识别了金黄色葡萄球菌细胞。还将其他几种常见于环境中的细菌,如大肠杆菌和枯草芽孢杆菌,与生物探针一起孵育,以测试其特异性。结果发现,新型 QDs-CMCS-IgG 生物探针对金黄色葡萄球菌细胞具有特异性识别能力。使用荧光分光光度计进行荧光测量可用于定量金黄色葡萄球菌细胞。在 600nm 处的样品荧光强度与细胞浓度在 10(3)-10(7) cfu/ml 的范围内成正比,检测限低至 900 cfu/ml。考虑到该方法的简单性和成本效益,预计其将在临床、食品和环境样品中用于鉴定和定量细菌。