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子系统。用4，4‘-联吡啶修饰和单独使用基面热解石墨电极进行了循环伏安测试。细胞与电极之间的电子传递是由存在于细胞壁的辅酶A介导的。根据循环伏安图的峰电流确定各种微生物细胞的数量。该系统将细菌分为革兰氏阳性菌和革兰氏阴性菌。革兰氏阳性菌的峰电流为0.65~0.69V，而饱和甘汞电极(SCE)为0.65~0.69V。革兰氏阴性杆菌的峰电位为0.70~0.74V，相对姐妹染色单体交换。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对革兰氏阴性和革兰氏阳性细菌进行分类。利用电极系统对野生和耐药细胞进行分类。微生物细胞在3…下培养在含有抗生素的培养液中7C反应1h以上，在0~1.0V范围内循环伏安曲线对姐妹染色单体交换有明显的抑制作用。野生株的峰电流降低或保持不变，而耐药株的峰电流增加。该系统还可用于尿路感染的诊断和抗生素的选择。在测量之前，通过将含有细菌的尿液与Rogosa培养液孵育3h来操纵活细胞数量。循环伏安曲线的峰电流随着尿液中大肠杆菌初始浓度的增加而增大。从细菌在尿液和乐果混合液中孵育3h的峰电流测定细菌对各种抗生素的敏感性。该系统检测T细胞、B细胞、巨噬细胞和肿瘤细胞等动物细胞。该系统还应用于结合抗原-抗体反应的细胞分类。因此，该系统在医疗和工业领域具有很好的适用性。较少

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