Direct identification of bacteria causing urinary tract infections by combining matrix-assisted laser desorption ionization-time of flight mass spectrometry with UF-1000i urine flow cytometry

Wang, X.-H.; Zhang, G.; Fan, Yongping; Yang, Xiu‐Wei; Sui, Wenjun; Lu, Xinxin

doi:10.1016/j.mimet.2012.12.016

Cited by 93 publications

(106 citation statements)

References 21 publications

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“…One limitation is the large amount of protein necessary to obtain reliable profiles. Therefore, as suggested by Wang et al (11), samples need to be preselected depending on the bacterial load. Another useful algorithm for the rapid diagnosis of UTIs is presented by Burillo et al (20), who combined Gram stain with mass spectrometry.…”

Section: Discussionmentioning

confidence: 99%

“…Matrix-assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF MS) has been suggested as a fast and reliable method for bacterial identification (8). This methodology has successfully been used for the rapid identification of microorganisms, both from culture plates and positive blood cultures, and also on other clinical samples, such as those from urine (9)(10)(11)(12)(13)(14). A biomarker capable of predicting the ability of MALDI-TOF MS to identify microorganisms directly from clinical samples would be useful to improve the diagnostic procedure, reducing costs and time.…”

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confidence: 99%

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

et al. 2016

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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 ionization-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 triplecentrifuged 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 microorganisms were misidentified, and no Candida spp. were correctly identified. Regarding the data from autoanalyzers, the best bacteriuria 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 containing Gram-negative bacteria. U rinary tract infections (UTIs) are among the most common nosocomial and community-acquired bacterial infections (1). The etiology is varied, but in approximately 90% of cases, enteric bacteria are implicated, especially Escherichia coli, which produces Ͼ70% of these infections. Other urinary tract pathogens are Klebsiella spp., Enterobacter spp., Proteus spp., Pseudomonas spp., Enterococcus spp., and Staphylococcus saprophyticus (2).A rapid diagnosis of UTIs has a significant beneficial impact on patient health, since it reduces unnecessary or inadequate empirical antimicrobial therapy (3). Urine culture is still the gold standard for the microbiological confirmation of UTIs; however, it takes 24 to 48 h to provide results. The use of screening methods, such as traditional Gram staining or other methods for counting urine particles, like flow cytometry or automated microscopic urine sediment analysis, allow the prompt prediction of negative samples and a preliminary identification of microorganisms in positive samples (4-7). Nevertheless, this information obtained from these methods is insufficient and requires culture and other biochemical tests.In recent years, proteomic techniques have achieved a relevant role in the identification of microorganisms in the field of clinical microbiology. Matrix-assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF MS) has been suggested as a...

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Section: Discussionmentioning

confidence: 99%

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

et al. 2016

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“…Urine flow cytometry may be used to screen out negative urines, with MALDI-TOF MS reserved to test the positive urine samples (55,56 ). Urine must be processed before MALDI-TOF MS testing.…”

Section: Direct Testing Of Clinical Samplesmentioning

confidence: 99%

MALDI-TOF MS for the Diagnosis of Infectious Diseases

2015

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BACKGROUND:First introduced into clinical microbiology laboratories in Europe, MALDI-TOF MS is being rapidly embraced by laboratories around the globe. Although it has multiple applications, its widespread adoption in clinical microbiology relates to its use as an inexpensive, easy, fast, and accurate method for identification of grown bacteria and fungi based on automated analysis of the mass distribution of bacterial proteins.CONTENT: This review provides a historical perspective on this new technology. Modern applications in the clinical microbiology laboratory are reviewed with a focus on the most recent publications in the field. Identification of aerobic and anaerobic bacteria, mycobacteria, and fungi are discussed, as are applications for testing urine and positive blood culture bottles. The strengths and limitations of MALDI-TOF MS applications in clinical microbiology are also addressed.

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“…Algunos estudios han evaluado el desempeño del MALDI-TOF MS para la identificación directa de microorganismos en muestras de orina, en conjunción con el uso de pruebas de tamización iniciales para determinar la positividad de la muestra. En el estudio de Wang y colaboradores 42 , se realizó la comparación entre los resultados de los métodos convencionales de urocultivo, con MALDI-TOF MS con un espectrómetro de masas Auto-flex III (Bruker Daltonics Inc.) acoplado a citometría de flujo (UF-1000i) para tamizar las orinas positivas de individuos con sintomatología de infección urinaria. Los resultados de MALDI-TOF MS fueron consistentes con los obtenidos por los métodos convencionales en 1373 de las 1456 muestras analizadas (94,3%), incluyendo 982 muestras negativas.…”

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La espectrometría de masas MALDI-TOF en el laboratorio de microbiología clínica

Maldonado

Robledo

2017

Infect

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ResumenLos métodos fenotípicos empleados para la identificación de microorganismos dependen de procesos metabólicos que requieren de tiempos de incubación mínimos para alcanzar resultados confiables. La espectrometría de masas MALDI-TOF (desorción/ionización láser asistida por una matriz con detección de masas por tiempo de vuelo) se ha instaurado como una metodología relevante para la identificación de microorganismos mediante el análisis de proteínas, a través de la creación de un espectro de masas específico de género y especie. En esta revisión, se presenta MALDI-TOF MS como una tecnología precisa para la identificación de bacterias, levaduras, mohos, en incluso de virus, que además, permite la reducción del tiempo para obtener un resultado de identificación, que puede impactar los costos de atención y duración de la estancia hospitalaria. La identificación de microorganismos directamente de muestras biológicas y la detección de mecanismos de resistencia a antimicrobianos, prometen un mayor impacto clínico y epidemiológico con el desarrollo e implementación de esta tecnología en los laboratorios de microbiología clínica.Palabras clave: MALDI-TOF MS, espectrometría de masas, diagnóstico microbiológico, identificación de patógenos, proteómica, infección del torrente sanguíneo, infección del tracto urinario. MALDI-TOF mass spectrometry in clinical microbiology laboratories. AbstractPhenotypic methods used for the identification of microorganisms depend on metabolic processes that require minimum incubation times to achieve reliable results. For this reason, MALDI-TOF MS (Matrix Assisted Laser Desorption/Ionization Mass Spectrometry) has been established as a relevant methodology for the identification of microorganisms using analysis of proteins, through the creation of a mass spectrum specific for genus and species. In the present review, MALDI-TOF MS is presented as an accurate technology for identifying bacteria, yeasts, molds and viruses; Its use allows reduction of the time to obtain an identification result, which may impact the costs of care and length of hospital stay. The identification of microorganisms directly from biological samples and the detection of mechanisms of antimicrobial resistance, promise an additional clinical and epidemiological impact with the development and implementation of this technology in clinical microbiology laboratories.Keywords: MALDI-TOF MS, mass spectrometry, microbiological diagnostic, pathogen identification, proteomics, blood stream infection, urinary tract infection. REVISIÓN IntroducciónLa identificación de microorganismos se ha realizado tradicionalmente por métodos basados en tinciones que permiten la clasificación de la morfología microscópica con el fin de apoyar decisiones diagnósticas y terapéuticas tempranas; así como en pruebas in vitro basadas en reacciones bioquímicas utilizadas por sistemas manuales o automatizados, que integran pruebas con diferentes sustratos, incrementando la rapidez y simplicidad de la identificación. Sin embargo, estos métodos fenotí...

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Direct identification of bacteria causing urinary tract infections by combining matrix-assisted laser desorption ionization-time of flight mass spectrometry with UF-1000i urine flow cytometry

Cited by 93 publications

References 21 publications

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

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

MALDI-TOF MS for the Diagnosis of Infectious Diseases

La espectrometría de masas MALDI-TOF en el laboratorio de microbiología clínica

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