Identification of fungal microorganisms by MALDI-TOF mass spectrometry

Chalupová, Jana; Raus, Martin; Sedlářová, Michaela; Šebela, Marek

doi:10.1016/j.biotechadv.2013.11.002

Cited by 159 publications

(108 citation statements)

References 91 publications

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“…This technique requires slight Mass Spectral (MS) ability, the utilize of uncomplicated instruments and relatively fast operator training. After comparing mass spectral identification with DNA analysis based technologies, it was found that MALDI-TOF MS requires smaller amounts of biological material and involves simpler sample preparation protocols with no initial assessment, such as Gram staining (Ilina et al, 2009;Chalupová et al, 2014;Tomazia et al, 2014). In the current study, the performance of MALDI-TOF-MS was evaluated comparing with conventional methods for bacterial screening of clinical and subclinical mastitis in dairy cows.…”

Section: Introductionmentioning

confidence: 99%

Phenotypical and Mass Spectral Assessment Methods for Identification of Some Contagious Mastitis Pathogens

Behiry¹,

Zahran²,

Marzouk³

et al. 2014

American Journal of Microbiology

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Mastitis is one of the most economic disease affecting dairy cows worldwide. Identification of mastitis pathogens still depends principally on culture and phenotypical method, which is a difficult and timeconsuming. Newly, microbiologists have focused their attention on the use of Mass Spectrometry (MS) for microbial identification, especially Matrix Assisted Laser Desorption Ionization Time-Of-Flight (MALDI-TOF). Therefore, this study was designated to evaluate the ability of MALDI-TOF to identify some contagious mastitis pathogens comparing with phenotypical methods such as API panels and VITEK 2 system. A total of one hundred twenty of Staphylococcus aureus (S. aureus), Coagulase Negative Staphylococci (CNS) and Streptococcus agalactiae (Strept. agalactiae) strains isolated from milk of cows affected by clinical and subclinical mastitis were used in the study. According to the results, ~95% of S. aureus, 100% of CNS and Strept. agalactiae were correctly identified by MALDI TOF MS. All S. aureus isolates were then confirmed by a nuc-based PCR technique. While ~92% of S. aureus, 87% of Strept. agalactiae and 76% of CNS were identified by VITEK 2 system. Moreover, ~89% of S. aureus, 80% of Strept. agalactiae and 72% of CNS were identified by API system. In brief, the results demonstrated that MALDI-TOF is a fast and truthful technique which has the capability to replace conventional identification of several bacterial strains usually isolated in clinical laboratories of microbiology. Therefore, it is recomended that MALDI-TOF MS technology can be regularly used in veterinary laboratories for identification of different species of bacteria, particularly when failure of phenotypic methods forces clinical microbiologists.

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Section: Introductionmentioning

confidence: 99%

Phenotypical and Mass Spectral Assessment Methods for Identification of Some Contagious Mastitis Pathogens

Behiry¹,

Zahran²,

Marzouk³

et al. 2014

American Journal of Microbiology

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“…Recently, the MALDI-TOF MS approach was also applied to mold identification and was shown to be able to accurately identify a wide array of species from various genera or groups of clinical interest, including Aspergillus spp. and dermatophytes (33)(34)(35)(36)(37)(38)(39)(40)(41)(42)(43)(44). Species identification by MALDI-TOF MS within the Fusarium genus has not yet been thoroughly investigated.…”

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confidence: 99%

Use of Matrix-Assisted Laser Desorption Ionization–Time of Flight Mass Spectrometry for Identification of Molds of the Fusarium Genus

et al. 2015

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eThe rates of infection with Fusarium molds are increasing, and a diverse number of Fusarium spp. belonging to different species complexes can cause infection. Conventional species identification in the clinical laboratory is time-consuming and prone to errors. We therefore evaluated whether matrix-assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF MS) is a useful alternative. The 289 Fusarium strains from the Belgian Coordinated Collections of Microorganisms (BCCM)/Institute of Hygiene and Epidemiology Mycology (IHEM) culture collection with validated sequence-based identities and comprising 40 species were used in this study. An identification strategy was developed, applying a standardized MALDI-TOF MS assay and an in-house reference spectrum database. In vitro antifungal testing was performed to assess important differences in susceptibility between clinically relevant species/species complexes. We observed that no incorrect species complex identifications were made by MALDI-TOF MS, and 82.8% of the identifications were correct to the species level. This success rate was increased to 91% by lowering the cutoff for identification. Although the identification of the correct species complex member was not always guaranteed, antifungal susceptibility testing showed that discriminating between Fusarium species complexes can be important for treatment but is not necessarily required between members of a species complex. With this perspective, some Fusarium species complexes with closely related members can be considered as a whole, increasing the success rate of correct identifications to 97%. The application of our user-friendly MALDI-TOF MS identification approach resulted in a dramatic improvement in both time and accuracy compared to identification with the conventional method. A proof of principle of our MALDI-TOF MS approach in the clinical setting using recently isolated Fusarium strains demonstrated its validity.

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“…In contrast, of the 29 isolates that saw increased scores with the analysis of younger colonies, 7 (24%) changed from incorrect to correct identifications. The reason for improved scores and identification at earlier stages of growth is unclear, but the improvement may be due to a reduced influence on the spectra of secondary characteristics, such as pigments and aerial mycelia, that may vary with time and by isolate (11). As these data were obtained by routine ethanol-formic acidacetonitrile extraction (7), it is evident that testing of isolates by standard sample preparation methods, but at earlier stages of growth, can improve the effectiveness of Nocardia isolate identification to the species level by MALDI-TOF MS.…”

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confidence: 99%

Identification of Nocardia Species by Matrix-Assisted Laser Desorption Ionization–Time of Flight Mass Spectrometry

et al. 2015

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b Matrix-assisted laser desorption ionization-time of flight mass spectrometry for identification of Nocardia species remains challenging. By identifying 83.1% (64 of 77) and 80% (8 of 10) to the species and complex levels, respectively, and 94.3% (82 of 87) to the genus level, we show that an approach using routine sample preparation, an up-to-date commercial database minimally augmented with custom spectra, and testing at an early stage of growth is promising. N ocardia species are aerobic actinomycetes belonging to the family Corynebacteriaceae. They are Gram-positive, weakly acid-fast environmental saprophytes with diverse colony morphologies and are the most commonly isolated aerobic actinomycete human pathogens (1). Nocardia infections generally result either from trauma-related introduction of the organism or from inhalation, particularly in immunocompromised patients. Pulmonary nocardiosis is characterized by pneumonia and can progress to a cavitary disease that may resemble tuberculosis. Disseminated disease, central nervous system involvement, and indolent pulmonary disease with cavities or contiguous spread are among the indicators used to test for Nocardia species (1, 2).Identification of Nocardia species is often performed by 16S rRNA gene sequencing (1, 3, 4), but matrix-assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF MS)-based identification has the potential to be a rapid and inexpensive alternative (5). Unfortunately, routine MALDI-TOF MS for identification of Nocardia species has proven difficult (6-8). Previous studies have stressed the need for enhanced sample preparation methods and/or considerably augmented reference spectrum databases to sufficiently identify Nocardia spp. (6,8). In this study, we demonstrated that the age of Nocardia cultures plays an important role in the success of MALDI-TOF MS identification. In addition, we showed that additional extraction steps beyond those recommended by the manufacturer are not required and that relatively modest augmentation of the current database is sufficient for the identification of routinely encountered Nocardia spp. By optimizing testing conditions using these approaches, we were able to improve the performance of MALDI-TOF MS for identification of Nocardia species.Clinical isolates (n ϭ 79) and type strains (n ϭ 8) of Nocardia species, selected on the basis of the frequency and diversity of isolates identified at ARUP Laboratories, were retrospectively tested by MALDI-TOF MS (Bruker Daltonics). These 87 isolates represented 25 unique Nocardia species identified to the species (n ϭ 77) or complex (n ϭ 10) level by partial 16S rRNA gene sequencing (Table 1). Reference spectra were created from the following additional 13 isolates identified to the species level by sequencing multiple genes (16S rRNA gene, hsp65, and secA1) (3, 9, 10): N. abscessus, N. araoensis, N. asiatica, N. asteroides, N. beijingensis, N. blacklockiae, N. brasiliensis, N. brevicatena, N. pseudobrasiliensis, N. puris, N. transvalensis, N...

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Identification of fungal microorganisms by MALDI-TOF mass spectrometry

Cited by 159 publications

References 91 publications

Phenotypical and Mass Spectral Assessment Methods for Identification of Some Contagious Mastitis Pathogens

Phenotypical and Mass Spectral Assessment Methods for Identification of Some Contagious Mastitis Pathogens

Use of Matrix-Assisted Laser Desorption Ionization–Time of Flight Mass Spectrometry for Identification of Molds of the Fusarium Genus

Identification of Nocardia Species by Matrix-Assisted Laser Desorption Ionization–Time of Flight Mass Spectrometry

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