Pharmazeutische Biotechnologie, Institut für Pharmazie, Ernst-Moritz-Arndt-Universität, 17487 Greifswald, Germany.
Appl Microbiol Biotechnol. 2010 Feb;85(5):1619-27. doi: 10.1007/s00253-009-2347-3. Epub 2009 Dec 2.
A new approach for the detection of virulence factors of Staphylococcus aureus and Staphylococcus epidermidis using an electrical protein array chip technology is presented. The procedure is based on an enzyme-linked sandwich immunoassay, which includes recognition and binding of virulence factors by specific capture and detection antibodies. Detection of antibody-bound virulence factors is achieved by measuring the electrical current generated by redox recycling of an enzymatically released substance. The current (measured in nanoampere) corresponds to the amount of the target molecule in the analyzed sample. The electrical protein chip allows for a fast detection of Staphylococcus enterotoxin B (SEB) of S. aureus and immunodominant antigen A homologue (IsaA homologue) of S. epidermidis in different liquid matrices. The S. aureus SEB virulence factor could be detected in minimal medium, milk, and urine in a concentration of 1 ng/ml within less than 23 min. Furthermore, a simultaneous detection of SEB of S. aureus and IsaA homologue of S. epidermidis in a single assay could be demonstrated.
提出了一种使用电蛋白芯片技术检测金黄色葡萄球菌和表皮葡萄球菌毒力因子的新方法。该方法基于酶联夹心免疫测定法，包括通过特异性捕获和检测抗体识别和结合毒力因子。通过测量酶释放物质的氧化还原循环产生的电流来检测与抗体结合的毒力因子。电流（以纳安为单位测量）与分析样品中目标分子的量相对应。电蛋白芯片允许快速检测金黄色葡萄球菌肠毒素 B (SEB) 和表皮葡萄球菌免疫显性抗原 A 同源物 (IsaA 同源物) 在不同液体基质中的存在。金黄色葡萄球菌 SEB 毒力因子可在少于 23 分钟内在最小培养基、牛奶和尿液中以 1 ng/ml 的浓度检测到。此外，还可以在单个测定中同时检测金黄色葡萄球菌的 SEB 和表皮葡萄球菌的 IsaA 同源物。
