Haiko Johanna, Savolainen Laura E, Hilla Risto, Pätäri-Sampo Anu
Department of Clinical Microbiology, University of Helsinki and Helsinki University Hospital, HUSLAB, Helsinki, Finland.
J Microbiol Methods. 2016 Oct;129:81-84. doi: 10.1016/j.mimet.2016.08.006. Epub 2016 Aug 5.
Complicated urinary tract infections, such as pyelonephritis, may lead to sepsis. Rapid diagnosis is needed to identify the causative urinary pathogen and to verify the appropriate empirical antimicrobial therapy. We describe here a rapid identification method for urinary pathogens: urine is incubated on chocolate agar for 3h at 35°C with 5% CO2 and subjected to MALDI-TOF MS analysis by VITEK MS. Overall 207 screened clinical urine samples were tested in parallel with conventional urine culture. The method, called U-si-MALDI-TOF (urine short incubation MALDI-TOF), showed correct identification for 86% of Gram-negative urinary tract pathogens (Escherichia coli, Klebsiella pneumoniae, and other Enterobacteriaceae), when present at >10(5)cfu/ml in culture (n=107), compared with conventional culture method. However, Gram-positive bacteria (n=28) were not successfully identified by U-si-MALDI-TOF. This method is especially suitable for rapid identification of E. coli, the most common cause of urinary tract infections and urosepsis. Turnaround time for identification using U-si-MALDI-TOF compared with conventional urine culture was improved from 24h to 4-6h.
复杂的尿路感染,如肾盂肾炎,可能会导致败血症。需要快速诊断以确定引起感染的尿路病原体,并验证适当的经验性抗菌治疗。我们在此描述一种尿路病原体的快速鉴定方法:将尿液在巧克力琼脂上于35°C、5%二氧化碳环境下孵育3小时,然后通过VITEK MS进行基质辅助激光解吸电离飞行时间质谱(MALDI-TOF MS)分析。总共207份经筛选的临床尿液样本与传统尿液培养法同时进行检测。这种方法称为U-si-MALDI-TOF(尿液短孵育MALDI-TOF),与传统培养方法相比,当培养物中革兰氏阴性尿路病原体(大肠杆菌、肺炎克雷伯菌和其他肠杆菌科细菌)浓度>10⁵cfu/ml时(n = 107),其对86%的此类病原体鉴定正确。然而,U-si-MALDI-TOF未能成功鉴定革兰氏阳性菌(n = 28)。该方法特别适用于快速鉴定尿路感染和尿脓毒症最常见的病因——大肠杆菌。与传统尿液培养相比,使用U-si-MALDI-TOF进行鉴定的周转时间从24小时缩短至4 - 6小时。