对数期金黄色葡萄球菌的离子交换色谱行为及其表征

Ion-Exchange Chromatographic Behavior and Characterization of Staphylococcus aureus in Exponential Phase

建立了高效离子交换色谱和激光光散射仪联用的分离 检测系统研究对数期金黄色葡萄球菌(Staphylococcusaureus)的方法。实验采用TSKgelSuperQ TOYOPEARL650C色谱柱,以20mmol/L的哌嗪 盐酸缓冲液(pH6 0)为流动相,倍增系数为21的光散射系统检测,发现对数期的S.aureus表现出两个有规律变化的色谱峰。透射电镜观察显示前峰的S.aureus为圆形,后峰为椭圆形且有一条明显的横隔壁。采用3 (4,5) 双甲基 2 噻唑 (2,5) 苯基溴化四氮唑盐(MTT)试剂测定这两个色谱峰,发现后峰的增殖能力比前峰高65 3%。实验结果表明,前峰为停止分裂的金黄色葡萄球菌、后峰为处于裂殖状态的金黄色葡萄球菌。该方法可作为研究微生物的一种快速而精确的新型分析方法。

A rapid, simple and sensitive method was developed to characterize S.aureus in exponential phase by a high performance ion-exchange chromatograph equipped with a laser light- scattering detector. An ion-exchange column (TSKgel SuperQ-TOYOPEARL 650C column, 7.6 mm i.d.×100 mm, Tosoh, Japan) was used for the separation. The mobile phase was a 20 (mmol/L) piperazine-hydrochloric acid (pH 6.0) buffer. It was found that S.aureus in exponential phase exhibited two elution peaks. Transmission electron microscope (TEM) showed that the bacterial cells in the first peak appeared to be circular, while the cells in the second one appeared to be elliptical and had one obvious diaphragm. It is found that the proliferation capability of the second fraction was better than the first fraction by 65.3%. These results suggested that the first fraction was the S.aureus that stopped dividing, while the second one was the S.aureus that were undergoing cell division. Therefore, the chromatographic method developed here may be a promising tool for the rapid and efficient separation and characterization of bacterial cells.