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

Sanguinetti, Maurizio

doi:10.1128/jcm.01640-16

Cited by 126 publications

(83 citation statements)

References 34 publications

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“…MALDI‐TOF MS is used in the analysis of portions of mycelium and/or conidia from fungal cultures, and is considered to be an accurate and rapid identification method . The aim of the present study was to evaluate the application of MALDI‐TOF‐MS for the identification of and discrimination between A flavus and A oryzae in clinical and environmental samples and compare its diagnostic power with that of β‐tubulin gene sequencing.…”

Section: Introductionmentioning

confidence: 99%

Discrimination of Aspergillus flavus from Aspergillus oryzae by matrix‐assisted laser desorption/ionisation time‐of‐flight (MALDI‐TOF) mass spectrometry

Hedayati

Armaki

Zarrinfar

et al. 2019

Mycoses

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Summary Background Aspergillus flavus is a major cause of severe non‐invasive fungal infections in the Middle Eastern countries. However, it is difficult to distinguish A flavus from A oryzae. Objectives To assess the potential of matrix‐assisted laser desorption/ionisation time‐of‐flight mass spectrometry (MALDI‐TOF MS) in discriminating between A flavus and A oryzae and compare it with β‐tubulin gene sequencing. Methods We used the Bruker Daltonik MALDI‐TOF MS system to analyse 200 clinical and environmental A flavus isolates and one A pseudonomius and one A alliaceus (Aspergillus section Flavi) isolate a priori identified as such by sequencing of the β‐tubulin gene. Results All 200 A flavus isolates were identified at the genus level and 176 (88%) at the species levels by MALDI‐TOF MS based on the spectral log‐scores (≥2.0 and 1.7‐1.99, respectively); among them, only 18 (10.2%) were confirmed as A flavus, whereas 35 (19.9%) were identified as A oryzae and 123 (69.9%) as A flavus/A oryzae. Aspergillus pseudonomius and A alliaceus were misidentified as A flavus and A parasiticus with log‐score values of 1.39 and 1.09, respectively. Conclusions The results indicate that the commercially available Bruker Daltonik MALDI‐TOF MS score database cannot separate A flavus and A oryzae species. We also showed that establishment of an in‐house library is a useful tool to discriminate closely related Aspergillus species, including A flavus and A oryzae.

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Section: Introductionmentioning

confidence: 99%

Discrimination of Aspergillus flavus from Aspergillus oryzae by matrix‐assisted laser desorption/ionisation time‐of‐flight (MALDI‐TOF) mass spectrometry

Hedayati

Armaki

Zarrinfar

et al. 2019

Mycoses

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“…For many years, the gold standard for diagnosis of superficial fungal infections has been direct microscopy often performed by the clinician, and culture sometimes supplemented by histopathology. Since the first dermatophyte polymerase chain reaction (PCR) was developed in 1996 an array of new molecular based methods for fungal identification have been developed with different PCR techniques and lately, matrix assisted laser desorption/ionization – time of flight (MALDI‐TOF) mass spectrometry has also been used . Because special equipment is required to perform molecular diagnostics the analysis is performed at specialized mycological or microbiological laboratories.…”

Section: Introductionmentioning

confidence: 99%

A survey among dermatologists: diagnostics of superficial fungal infections – what is used and what is needed to initiate therapy and assess efficacy?

Saunte

Piraccini

Sergeev

et al. 2018

Acad Dermatol Venereol

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Background Superficial fungal infections are common. It is important to confirm the clinical diagnosis by mycological laboratory methods before initiating systemic antifungal treatment, especially as antifungal sensitivity and in vitro susceptibility may differ between different genera and species. For many years, the gold standard for diagnosis of superficial fungal infections has been direct fungal detection in the clinical specimen (microscopy) supplemented by culturing. Lately, newer molecular based methods for fungal identification have been developed. Objective This study was initiated to focus on the current usage of mycological diagnostics for superficial fungal infections by dermatologists. It was designed to investigate whether it was necessary to differentiate between initial diagnostic tests and those used at treatment follow‐up in specific superficial fungal infections. Methods An online questionnaire was distributed among members of the EADV mycology Task Force and other dermatologists with a special interest in mycology and nail disease. Results The survey was distributed to 62 dermatologists of whom 38 (61%) completed the whole survey, 7 (11%) partially completed and 17 (27%) did not respond. Nearly, all respondents (82–100%) said that ideally they would use the result of direct microscopy (or histology) combined with a genus/species directed treatment of onychomycosis, dermatophytosis, Candida‐ and Malassezia‐related infections. The majority of the dermatologists used a combination of clinical assessment and direct microscopy for treatment assessment and the viability of the fungus was considered more important at this visit than when initiating the treatment. Molecular based methods were not available for all responders. Conclusion The available diagnostic methods are heterogeneous and their usage differs between different practices as well as between countries. The survey confirmed that dermatologists find it important to make a mycological diagnosis, particularly prior to starting oral antifungal treatment in order to confirm the diagnose and target the therapy according to genus and species.

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“…A variety of previous reports have highlighted the importance of supplementing the commercial databases supplied with the various MALDI-TOF platforms with appropriate reference MALDI-TOF spectra in order to improve identification success (29)(30)(31)(32). Since none of the 10 known species included in the test panel employed here were represented in the latest Bruker MSP database, appropriate reference spectra were generated and added to the commercial database.…”

Section: Discussionmentioning

confidence: 99%

“…Indeed, we recently demonstrated that MALDI-TOF MS could reliably discriminate between a wide variety of clinical and environmental Exophiala species (32). However, in all cases identification accuracy is limited by the availability of appropriate reference MALDI-TOF spectra in the commercial databases supplied with the various MALDI-TOF platforms (29)(30)(31)(32). Here we evaluated whether MALDI-TOF MS could be used to differentiate a variety of filamentous fungi implicated in black-grain eumycetoma and related subcutaneous clinical syndromes.…”

mentioning

confidence: 99%

“…Matrix-assisted laser desorption ionization-time of flight (MALDI-TOF) mass spectrometry (MS) has been shown to provide a robust, cost-effective, and rapid means of identification of a variety of clinical microorganisms from pure culture (reviewed in reference 29), including the majority of clinically relevant Candida and other yeast species (30) and some molds (31). Indeed, we recently demonstrated that MALDI-TOF MS could reliably discriminate between a wide variety of clinical and environmental Exophiala species (32).…”

mentioning

confidence: 99%

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Rapid and Robust Identification of the Agents of Black-Grain Mycetoma by Matrix-Assisted Laser Desorption Ionization–Time of Flight Mass Spectrometry

2017

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Eumycetoma, a chronic fungal infection endemic in India, Indonesia, and parts of Africa and South and Central America, follows traumatic implantation of saprophytic fungi and frequently requires radical surgery or amputation in the absence of appropriate treatment. Fungal species that can cause black-grain mycetomas include Madurella spp., Falciformispora spp., Trematosphaeria grisea, Nigrograna mackinnonii, Pseudochaetosphaeronema larense, Medicopsis romeroi, and Emarellia spp. Rhytidhysteron rufulum and Parathyridaria percutanea cause similar subcutaneous infections, but these infections lack the draining sinuses and fungal grains characteristic of eumycetoma. Accurate identification of the agents of subcutaneous fungal infection is essential to guide appropriate antifungal therapy. Since phenotypic identification of the causative fungi is often difficult, time-consuming molecular approaches are currently required. In the study described here we evaluated whether matrix-assisted laser desorption ionization-time of flight (MALDI-TOF) mass spectrometry might allow the accurate identification of eumycetoma agents and related fungi. A panel of 57 organisms corresponding to 10 different species from confirmed cases of eumycetoma and subcutaneous pedal masses, previously formally identified by PCR amplification and sequencing of internal transcribed spacer 1 (ITS1), was employed. Representative isolates of each species were used to create reference MALDI-TOF spectra, which were then used for the identification of the remaining isolates in a user-blinded manner. Here, we demonstrate that MALDI-TOF mass spectrometry accurately identified all of the test isolates, with 100%, 90.4%, and 67.3% of isolates achieving log scores greater than 1.8, 1.9, and 2.0, respectively. KEYWORDS black-grain mycetoma, Madurella, Pleosporales, Trematosphaeriaceae, Emarellia, Biatriospora, Rhytidhysteron, Medicopsis, Falciformispora, Exophiala, MALDI-TOF, Pleosporales B lack-grain eumycetoma is an ancient, debilitating fungal infection that is endemic in India, Indonesia, and parts of Africa and South America (1). Disease progression is protracted and results from the traumatic implantation of pigmented saprophytic fungi, usually on exposed body sites. The disease is characterized by a chronic progressive destruction of soft tissue and adjoining structures with associated tumefaction and the formation of purosanguineous sinuses that drain fungal grains (2-5). Without appropriate treatment, the infection eventually progresses to involve the skeletal system, and clinical and mycological cure is unlikely without extensive debridement/ amputation (2-5). The extruded grains are either pale or dark (black), depending on the etiological agent (4, 5). The principal etiological agents described to date include Madurella spp. (Sordariales); a host of fungi classified in the Pleosporales, including

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

Cited by 126 publications

References 34 publications

Discrimination of Aspergillus flavus from Aspergillus oryzae by matrix‐assisted laser desorption/ionisation time‐of‐flight (MALDI‐TOF) mass spectrometry

Discrimination of Aspergillus flavus from Aspergillus oryzae by matrix‐assisted laser desorption/ionisation time‐of‐flight (MALDI‐TOF) mass spectrometry

A survey among dermatologists: diagnostics of superficial fungal infections – what is used and what is needed to initiate therapy and assess efficacy?

Rapid and Robust Identification of the Agents of Black-Grain Mycetoma by Matrix-Assisted Laser Desorption Ionization–Time of Flight Mass Spectrometry

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