Basic studies on development of the automatic cell detection system using voltammetry

开发了用于微生物细胞和动物细胞的检测和分类的电极系统。用单独使用或用4，4 ′-联吡啶修饰的基面热解石墨电极获得循环伏安图。细胞和电极之间的电子传递由存在于细胞壁中的辅酶A介导。根据循环伏安图的峰电流确定各种微生物细胞的数量。肠道菌群分为革兰氏阳性菌和革兰氏阴性菌。革兰氏阳性菌在饱和甘汞电极（SCE）上的峰电流为0.65 ~ 0.69V。革兰氏阴性菌的峰电位为0.70 ~ 0.74Vvs SCE。根据重复两次电位扫描时第二峰电流与第一峰电流的比值将革兰氏阴性菌和革兰氏阳性菌进行了分类，并利用该电极系统对野生菌和抗生素耐药菌进行了分类。微生物细胞在3 ℃下孵育 ...更多信息 在0 ~ 1.0V（vs. SCE）范围内获得了循环伏安图。野生型菌株的峰电流降低或不变，而耐药菌株的峰电流升高。该系统也可用于尿路感染的诊断和抗生素筛选。在测量之前，通过将含有细菌的尿液与Rogosa培养基孵育3小时来操纵活细胞数。循环伏安峰电流随尿液中大肠杆菌初始浓度的增加而增大。用该系统检测了T细胞、B细胞、巨噬细胞和肿瘤细胞等动物细胞。该系统也被应用到细胞分类与抗原抗体反应耦合。因此，该系统被证明适用于医疗和工业领域。少

Electrodesystems were developed for the detection and classification of microbial cells and animal cells. Cyclic voltammograms were obtained with a basal plane pyrolytic graphite electrode used alone or modified with 4,4'-bipyridine. Electron transfer between cells and the electrode was mediated by coenzyme A present in the cell wall. The number of various microbial cells was determined from the peak current of cyclic voltammograms. Intenstinal bacteria were classified as gram-positive or gram-negative by the system. Gram-positive bacteria gave peak currents at 0.65 to 0.69V versus the saturated calomel electrode (SCE). The peak potentials of gram-negative bacteria were 0.70 to 0.74V versus SCE. Gram-negative and gram-positive bacteria were also classified based on the ratio of the second peak current to be first peak current when the potential sweep was repeated twice.Wild and antibiotic resistant cells were also classified with the electrode system. Microbial cells were incubted at 3 … More 7 C for 1h in the medium containing antibiotics, and cyclic voltammograms were obtained in the range of 0 to 1.0V versus SCE. The peak current of wild strains decreased or constant whereas the peak current of antibiotic resistant strains increased.The system was also applied to diagnosis of urinary tract infection and antibiotic selection. Before measurements, viable cell numbers were manipulated by incubating urine containing bacteria with Rogosa medium for 3h. The peak current of cyclic voltammogram increased with increasing the initial cell concentration of E.coli in urine. Susceptability of bacteria to various antibiotics was also determined from the peak current when they were incubated for 3h in the mixture of urine and Rogosa medium.Animal cells such as T cells, B cells, macrophage and tumor cells were detected by the system. The system was also applied to the cell classification coupled with antigen-antibody reactions. Thus, the system was shown to be applicable in the medical and industrial fields. Less