Characterization of intact <i>Penicillium</i> spores by matrix‐assisted laser desorption/ionization mass spectrometry

Chen, Hun Yun; Chen, Yu-Chie

doi:10.1002/rcm.2229

Cited by 74 publications

(48 citation statements)

References 32 publications

(36 reference statements)

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“…The technique allows rapid and reliable identification of bacterial taxa (Bright et al 2002;Dickinson et al 2004;Fenselau and Demirev 2001;Friedrichs et al 2007;Hathout et al 1999;Lasch et al 2008;Pignone et al 2006;Ryzhov et al 2000;Smole et al 2002) and has already been successfully applied to the identification of specimens belonging to the fungal genera Serpula, Conidiophora, Antrodia, Penicillium, and Candida (Chen and Chen 2005;Qian et al 2008;Schmidt and Kallow 2005). Recently, MALDI-TOF MS has also been used to study the peptaibiomics (Degenkolb and Brückner 2008;Degenkolb et al 2008a, b;Neuhof et al 2007a;Toniolo and Brückner 2009) and for direct identification of hydrophobins of Trichoderma (Neuhof et al 2007b).…”

Section: Introductionmentioning

confidence: 97%

MALDI-TOF MS of Trichoderma: a model system for the identification of microfungi

Respinis

Vogel²,

Benagli

et al. 2009

Mycol Progress

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This investigation aimed to assess whether MALDI-TOF MS analysis of the proteome could be applied to the study of Trichoderma, a fungal genus selected because it includes many species and is phylogenetically well defined. We also investigated whether MALDI-TOF MS analysis of peptide mass fingerprints would reveal apomorphies that could be useful in diagnosing species in this genus. One hundred and twenty nine morphologically and genetically well-characterized strains of Hypocrea and Trichoderma, belonging to 25 species in 8 phylogenetic clades, were analyzed by MALDI-TOF MS mass spectrometry. The resulting peak lists of individual samples were submitted to single-linkage cluster analysis to produce a taxonomic tree and were compared to ITS and tef1 sequences from GenBank. SuperSpectra™ for the 13 most relevant species of Trichoderma were computed. The results confirmed roughly previously defined clades and sections. With the exceptions of T. saturnisporum (Longibrachiatum Clade) and T. harzianum (Harzianum Clade), strains of individual species clustered very closely. T. polysporum clustered distantly from all other groups. The MALDI-TOF MS analysis accurately reflected the phylogenetic classification reported in recent publications, and, in most cases, strains identified by DNA sequence analysis clustered together by MALDI-TOF MS. The resolution of MALDI-TOF MS, as performed here, was roughly equivalent to ITS rDNA. The MALDI-TOF MS technique analyzes peptides and represents a rough equivalent to sequencing, making this method a useful adjunct for determination of species limits. It also allows simple, reliable, and quick species identification, thus representing a valid alternative to gene sequencing for species diagnosis of Trichoderma and other fungal taxa.

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Section: Introductionmentioning

confidence: 97%

MALDI-TOF MS of Trichoderma: a model system for the identification of microfungi

Respinis

Vogel²,

Benagli

et al. 2009

Mycol Progress

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“…Furthermore, the fungal infection of economically important plants leads to contamination of human foodstuffs and animal feed causing poisoning of humans and breeding cattle with mycotoxins, toxic metabolites produced by many fungal species. [26][27][28][29] Previous studies [21][22][23][24] …”

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confidence: 98%

“…19,20 Only a few groups have focused on fungal cells, 21 in particular on fungal spores. [22][23][24] In contrast to bacteria, fungal cell walls are mainly composed of different polysaccharides (80-90%), including chitin which adds rigidity and structural stability to the cells, but peptides, proteins, lipids, polyphosphates and inorganic ions are also present. 25 The composition of fungal cell walls and also of fungal spores exhibits qualitative and quantitative differences within different fungal species, but also between different strains of the same fungal species, a prerequisite for the classification and differentiation of fungal species by MALDI-TOF MS.…”

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confidence: 99%

Evaluation of matrix‐assisted laser desorption/ionization (MALDI) preparation techniques for surface characterization of intact Fusarium spores by MALDI linear time‐of‐flight mass spectrometry

Kemptner

Marchetti‐Deschmann

Mach

et al. 2009

Rapid Comm Mass Spectrometry

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Unambiguous identification of mycotoxin-producing fungal species as Fusarium is of great relevance to agriculture and the food-producing industry as well as in medicine. Protein profiles of intact fungal spores, such as Penicillium, Aspergillus and Trichoderma, derived from matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) were shown to provide a rapid and straightforward method for species identification and characterization. In this study, we applied this approach to five different Fusarium spp. strains which are known to affect the growth of different grain plants. To obtain a suitable MALDI matrix system and sample preparation method, thin-layer, dried-droplet and sandwich methods and several MALDI matrices, namely CHCA, DHB, FA, SA and THAP dissolved in various solvent mixtures (organic solvents such as ACN, MeOH, EtOH and iPrOH and for the aqueous phase water and 0.1% TFA), were evaluated in terms of mass spectrometric pattern and signal intensities. The most significant peptide/protein profiles were obtained with 10 mg ferulic acid (FA) in 1 mL ACN/0.1% TFA (7:3, v/v) used as matrix system. Mixing the spores with the matrix solution directly on the MALDI target (dried-droplet technique) resulted in an evenly distributed spores/matrix crystal layer, yielding highly reproducible peptide/protein profiles from the spore surfaces. Numerous abundant ions throughout the investigated m/z range (m/z 1500-15 000) could be detected. Differences in the obtained mass spectral patterns allowed the differentiation of spores of various Fusarium species.

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“…The assessment of MALDI-TOF for species-level identification of filamentous fungi, however, is not as extensive. Many initial fungal MALDI-TOF MS studies focused on isolate identification using purified fungal spores (3,13,15,28). Only more recently has this technology been applied directly to unfractionated fungal colonies (10,11,17).…”

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confidence: 99%

Dermatophyte Identification Using Matrix-Assisted Laser Desorption Ionization–Time of Flight Mass Spectrometry

Theel¹,

Hall²,

Mandrekar

et al. 2011

J Clin Microbiol

108

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The performance of the Bruker Biotyper matrix-assisted laser desorption ionization-time of flight (MALDI-TOF) mass spectrometer (MS) for the identification of dermatophytes from clinical cultures was compared to that of dermatophyte identification using 28S rRNA gene sequencing. The MALDI Biotyper library (MBL; version 3.0) was used alone and in combination with a supplemented library containing an additional 20 dermatophyte spectra (S-MBL). Acquired spectra were interpreted using both the manufacturer-recommended scores (genus, >1.7; species, >2.0) and adjusted cutoff values established by this study (genus, >1.5; species, >1.7); identifications required a minimum 10% difference in scores between the top two different organisms to be considered correct. One hundred well-characterized, archived dermatophyte isolates and 71 fresh dermatophyte cultures were evaluated using both libraries and both sets of cutoff criteria. Collectively, the S-MBL significantly outperformed the MBL at both the genus (93% versus 37.4%; P < 0,0001) and species (59.6% versus 20.5%; P < 0.0001) levels when using the adjusted score criteria. Importantly, application of the lowered cutoff values significantly improved genus (P ‫؍‬ 0.005)-and species (P < 0.0001)-level identification for the S-MBL, without leading to an increase in misidentifications. MALDI-TOF MS is a cost-effective and rapid alternative to traditional or molecular methods for dermatophyte identification, provided that the reference library is supplemented to sufficiently encompass clinically relevant, intraspecies strain diversity.Dermatophytes, comprising species within the genera Epidermophyton, Microsporum, and Trichophyton, exclusively invade and infect keratinized tissues (i.e., hair, skin, and nails). These organisms are among the most common fungal infections in humans, accounting for over 5 million physician visits annually in the United States (7, 26). Dermatophytes primarily cause superficial dermal mycoses and rarely progress to more invasive disease (8). In the United States, Trichophyton rubrum remains the most common cause of tinea infections and the most frequently isolated dermatophytic pathogen (7,24).Laboratory identification of dermatophytes is typically based on macroscopic observation of colony morphology (i.e., pigmentation, growth rate, texture, etc.) on selective media, followed by microscopic examination of conidia. In addition, specialized Trichophyton agars may be used for differentiation of species within the genera (14). These phenotypic techniques require experienced technologists and are often labor-intensive with a prolonged turnaround time. With the advancement of modern molecular techniques, some laboratories have turned to dermatophyte identification schemes involving sequencing of specific ribosomal DNA regions or real-time PCR amplification of discrete genes (2, 20). While these methods are highly accurate and rapid, they are costly and can be complex to implement.Matrix-assisted laser desorption ionization-time of flight (MALDI-TOF) m...

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Characterization of intact Penicillium spores by matrix‐assisted laser desorption/ionization mass spectrometry

Cited by 74 publications

References 32 publications

MALDI-TOF MS of Trichoderma: a model system for the identification of microfungi

MALDI-TOF MS of Trichoderma: a model system for the identification of microfungi

Evaluation of matrix‐assisted laser desorption/ionization (MALDI) preparation techniques for surface characterization of intact Fusarium spores by MALDI linear time‐of‐flight mass spectrometry

Dermatophyte Identification Using Matrix-Assisted Laser Desorption Ionization–Time of Flight Mass Spectrometry

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