Direct Identification of Urinary Tract Pathogens from Urine Samples, Combining Urine Screening Methods and Matrix-Assisted Laser Desorption Ionization–Time of Flight Mass Spectrometry

Abstract

Early diagnosis of urinary tract infections (UTIs) is essential to avoid inadequate or unnecessary empirical antibiotic therapy. Microbiological confirmation takes 24 to 48 h. The use of screening methods, such as cytometry and automated microscopic

analysis of urine sediment, allows the rapid prediction of negative samples. In addition, matrix-assisted laser desorption ioniza- tion–time of flight mass spectrometry (MALDI-TOF MS) is a widely established technique in clinical microbiology laboratories

used to identify microorganisms. We evaluated the ability of MALDI-TOF MS to identify microorganisms from direct urine samples and the predictive value of automated analyzers for the identification of microorganisms in urine by MALDI-TOF MS. A total of 451 urine samples from patients with suspected UTIs were first analyzed using the Sysmex UF-1000i flow cytometer, an

automatic sediment analyzer with microscopy (SediMax), culture, and then processed by MALDI-TOF MS with a simple triple- centrifuged procedure to obtain a pellet that was washed and centrifuged and finally applied directly to the MALDI-TOF MS

plate. The organisms in 336 samples were correctly identified, mainly those with Gram-negative bacteria (86.10%). No microor- ganisms were misidentified, and no Candida spp. were correctly identified. Regarding the data from autoanalyzers, the best bac- teriuria cutoffs were 1,000 and 200 U/l for UF-1000i and SediMax, respectively. It was concluded that the combination of a

urine screening method and MALDI-TOF MS provided a reliable identification from urine samples, especially in those contain- ing Gram-negative bacteria.