Unidad de Investigación, Hospital Universitario de Salamanca, Salamanca, Spain.
J Clin Microbiol. 2010 Jun;48(6):2110-5. doi: 10.1128/JCM.02215-09. Epub 2010 Apr 14.
Matrix-assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF MS) has been suggested as a reliable method for bacterial identification from cultures. Direct analysis of clinical samples might increase the usefulness of this method, shortening the time for microorganism identification. We compared conventional methods for the diagnosis of urinary tract infections (UTIs) and identification of the urinary tract pathogens (automated screening, plate cultures, and identification based on biochemical characteristics) and a fast method based on conventional screening and MALDI-TOF MS. For this latter method, 4 ml of urine was centrifuged at a low-revolution setting (2,000 x g) to remove leukocytes and then at high revolutions (15,500 x g) to collect bacteria. The pellet was washed and then applied directly to the MALDI-TOF MS plate. Two hundred sixty urine samples, detected as positive by the screening device (UF-1000i), were processed by culture and MALDI-TOF MS. Twenty samples were positive in the screening device but negative in culture, and all of them were also negative by MALDI-TOF MS. Two-hundred thirty-five samples displayed significant growth of a single morphological type in culture. Two-hundred twenty of them showed bacterial growth of >10(5) CFU/ml. Microorganism identifications in this group were coincident at the species level in 202 cases (91.8%) and at the genus level in 204 cases (92.7%). The most frequent microorganism was Escherichia coli (173 isolates). MALDI-TOF MS identified this microorganism directly from the urine sample in 163 cases (94.2%). Our results show that MALDI-TOF MS allows bacterial identification directly from infected urine in a short time, with high accuracy, and especially when Gram-negative bacteria with high bacterial counts are involved.
基质辅助激光解吸电离飞行时间质谱(MALDI-TOF MS)已被提议作为一种从培养物中鉴定细菌的可靠方法。直接分析临床样本可能会增加这种方法的实用性,缩短微生物鉴定的时间。我们比较了尿路感染(UTI)的常规诊断方法和尿路感染病原体的鉴定方法(自动筛选、平板培养和基于生化特征的鉴定)以及基于常规筛选和 MALDI-TOF MS 的快速方法。对于后一种方法,将 4 毫升尿液在低转速(2000 x g)下离心以去除白细胞,然后在高转速(15500 x g)下收集细菌。沉淀用洗涤液洗涤后直接应用于 MALDI-TOF MS 板。用筛选装置(UF-1000i)检测为阳性的 260 份尿液样本进行培养和 MALDI-TOF MS 处理。筛选装置阳性但培养阴性的 20 个样本,均为 MALDI-TOF MS 阴性。235 份样本在培养中显示出单一形态类型的显著生长。其中 220 份样本的细菌生长量>10(5) CFU/ml。在该组中,202 例(91.8%)在种水平上和 204 例(92.7%)在属水平上的微生物鉴定结果一致。最常见的微生物是大肠埃希菌(173 株)。MALDI-TOF MS 直接从 163 例(94.2%)尿液样本中鉴定出这种微生物。我们的结果表明,MALDI-TOF MS 可以在短时间内直接从感染尿液中鉴定细菌,具有很高的准确性,尤其是当涉及高细菌计数的革兰氏阴性菌时。