School of Medical Laboratory, Tianjin Medical University, 1 Guang Dong Road, Tianjin 300203, PR China.
J Chromatogr B Analyt Technol Biomed Life Sci. 2013 Sep 15;935:32-5. doi: 10.1016/j.jchromb.2013.07.011. Epub 2013 Jul 25.
This study explored a method that can rapidly detect bacteria in urine samples for the auxiliary determination of urinary tract infections (UTIs). Urine samples from patients with UTIs (230 cases) were obtained using aseptic technique. The urine biochemical assay was then carried out using an automated urine analyzer for all the urine samples. Bacterial species were identified by a combination of bacterial culture technique, morphological observation and the BACT-IST microbial identification/susceptibility analysis system. The most common seven species of bacteria in the study included Escherichia coli, Klebsiella pneumoniae, Pseudomonas aeruginosa, Enterobacter cloacae, Enterococcus faecalis, Staphylococcus aureus and Staphylococcus epidermidis. Bacterial samples were suspended in sample buffer solutions and separated by the "three-plug-injection" method using capillary electrophoresis (CE). Each species of bacteria appeared as a bacterial peak. The mixture of the seven species also provided only one peak. Further analysis showed that the concentration limit for the "three-plug-injection" method is 10(6) colony forming units (CFU)/mL, and there is a good linear relationship between the peak height and bacterial concentration (R(2)=0.99). The effect of urine composition on CE results was also investigated. The results showed that urine composition, i.e., proteins, white blood cells (WBCs) and red blood cells (RBCs), affected the peak retention time but could not affect the separation of bacteria. The results demonstrated that the bacteria in urine samples can be detected within 10min by the "three-plug-injection" method using CE. The "three-plug-injection" method is therefore suitable for the rapid detection of organisms in clinical urine samples from UTIs.
本研究探索了一种可快速检测尿液样本中细菌的方法,用于辅助诊断尿路感染(UTIs)。采用无菌技术采集尿路感染患者(230 例)的尿液样本。然后使用自动尿液分析仪对所有尿液样本进行尿液生化分析。通过细菌培养技术、形态观察和 BACT-IST 微生物鉴定/药敏分析系统相结合来鉴定细菌种类。本研究中最常见的七种细菌包括大肠埃希菌、肺炎克雷伯菌、铜绿假单胞菌、阴沟肠杆菌、粪肠球菌、金黄色葡萄球菌和表皮葡萄球菌。细菌样本悬浮在样品缓冲溶液中,通过毛细管电泳(CE)的“三插件注射”方法分离。每种细菌都呈现出一个细菌峰。七种细菌的混合物也只提供一个峰。进一步分析表明,“三插件注射”方法的浓度极限为 10(6) 菌落形成单位(CFU)/mL,并且峰高与细菌浓度之间存在良好的线性关系(R(2)=0.99)。还研究了尿液成分对 CE 结果的影响。结果表明,尿液成分(即蛋白质、白细胞(WBCs)和红细胞(RBCs))会影响峰保留时间,但不会影响细菌的分离。结果表明,CE 采用“三插件注射”方法可在 10min 内检测尿液样本中的细菌。因此,“三插件注射”方法适用于快速检测尿路感染患者临床尿液样本中的生物体。