Gupta Ashish, Dwivedi Mayank, Mahdi Abbas Ali, Gowda G A Nagana, Khetrapal Chunni Lal, Bhandari Mahendra
Centre of Bio-Medical Magnetic Resonance, Department of Microbiology, Sanjay Gandhi Postgraduate Institute of Medical Sciences, Lucknow, India.
BJU Int. 2009 Jul;104(2):236-44. doi: 10.1111/j.1464-410X.2009.08448.x. Epub 2009 Feb 23.
To address the shortcomings of urine culture for the diagnosis of urinary tract infection (UTI), we used 1H-nuclear magnetic resonance (NMR) spectroscopy for identifying and quantifying Escherichia coli, Pseudomonas aeruginosa, Klebsiella pneumonia and Proteus mirabilis.
PATIENTS, SUBJECTS AND METHODS: Urine samples from patients with suspected UTI (617), healthy volunteers (50) and commercially available standard strains of E. coli, K. pneumonia, P. aeruginosa, Enterobacter, Acinobacter, Pr. mirabilis, Citrobacter frundii, Streptococcus saprophyticus and Enterococcus faecalis were assessed between 2003 and 2006. 1H-NMR spectra were recorded on a 400 MHz spectrophotometer; to quantify the bacteria we estimated the areas under the spectral peaks of the specific metabolic product compared with the known concentration of trimethyl silyl propionic acid. All urine specimens were cultured in addition to an assessment by NMR spectroscopy.
Preliminary urinary spectroscopy of the unprocessed samples showed peaks of nonspecific metabolites such as succinate, acetate, lactate and ethanol, indicating infected samples. Based on the results from processed samples, 93% (240/256) of E. coli, 92% (101/110) of K. pneumoniae, 93% (56/60) of P. aeruginosa and eight of 10 Pr. mirabilis could be diagnosed with NMR (numerator) and urine culture (denominator). The remaining samples were sterile and/or had a bacterial population of <10(3) colony-forming units (CFU)/mL. The NMR method diagnosed bacterial densities of >10(3) CFU.
The identification of the common uropathogens E. coli, K. pneumoniae, P. aeruginosa and Pr. mirabilis by NMR spectroscopy has a shorter reporting time and can be used to differentiate between infected, contaminated and sterile specimens.
为解决尿培养在诊断尿路感染(UTI)方面的不足,我们采用1H-核磁共振(NMR)光谱法来鉴定和定量大肠杆菌、铜绿假单胞菌、肺炎克雷伯菌和奇异变形杆菌。
患者、受试者与方法:对2003年至2006年间疑似UTI患者的尿液样本(617份)、健康志愿者的尿液样本(50份)以及市售的大肠杆菌、肺炎克雷伯菌、铜绿假单胞菌、肠杆菌、不动杆菌、奇异变形杆菌、弗氏柠檬酸杆菌、腐生葡萄球菌和粪肠球菌标准菌株进行评估。在400 MHz分光光度计上记录1H-NMR光谱;为了定量细菌,我们将特定代谢产物光谱峰下的面积与已知浓度的三甲基硅基丙酸进行比较。除了通过NMR光谱法进行评估外,所有尿液标本均进行培养。
未经处理样本的初步尿液光谱显示出琥珀酸盐、乙酸盐、乳酸盐和乙醇等非特异性代谢物的峰,表明样本受到感染。根据处理后样本的结果,93%(240/256)的大肠杆菌、92%(101/110)的肺炎克雷伯菌、93%(56/60)的铜绿假单胞菌以及10株奇异变形杆菌中的8株可通过NMR(分子)和尿培养(分母)进行诊断。其余样本无菌和/或细菌数量<10(3) 菌落形成单位(CFU)/mL。NMR方法诊断出细菌密度>10(3) CFU。
通过NMR光谱法鉴定常见尿路病原体大肠杆菌、肺炎克雷伯菌、铜绿假单胞菌和奇异变形杆菌的报告时间更短,可用于区分感染、污染和无菌标本。