Evaluation of matrix-assisted laser desorption ionization–time-of-flight mass spectrometry to differentiate between Candida albicans and Candida dubliniensis

doi:10.1016/j.diagmicrobio.2016.01.017

Diagnostic Microbiology and Infectious Disease

Volume 85, Issue 1, May 2016, Pages 73-76

https://doi.org/10.1016/j.diagmicrobio.2016.01.017 Get rights and content

Highlights

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Evaluation of a well-characterized strain set of Candida albicans and Candida dubliniensis showed that matrix-assisted laser desorption ionization–time-of-flight mass spectrometry (MALDI-TOF-MS), when using the direct formic acid (FA) extraction method, was able to accurately discriminate between C. albicans and C. dubliniensis.
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Utilization of this technology will help to define the true epidemiological prevalence of C. dubliniensis and expand knowledge of what role this species contributes to human infection.
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Direct FA method for processing Candida species for MALDI-TOF-MS analysis is as effective as a traditional ethanol extraction process.

Abstract

Matrix-assisted laser desorption ionization–time-of-flight mass spectrometry (MALDI-TOF-MS) analysis in conjunction with the direct formic acid (FA) sample processing method was evaluated for the ability to differentiate the closely related species of Candida albicans and Candida dubliniensis. The results showed that MALDI-TOF-MS, using the direct FA method, was reliable to differentiate between these species.

Section snippets

Acknowledgements

The authors would like to thank Jon Biehle at Creighton University Medical Center, Omaha, NE, USA, and Ana Gales, University of Iowa, Iowa City, IA, USA, for the gift of some strains utilized in this study.

References (16)

W.Y. Jamal et al.
Comparative evaluation of two matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) systems for the identification of clinically significant yeasts
Int J Infect Dis
(2014)
A. Bizzini et al.
Performance of matrix-assisted laser desorption ionization-time of flight mass spectrometry for identification of bacterial strains routinely isolated in a clinical microbiology laboratory
J Clin Microbiol
(2010)
R. Ells et al.
Arachidonic acid increases antifungal susceptibility of Candida albicans and Candida dubliniensis
J Antimicrob Chemother
(2009)
J. Gutierrez et al.
Candida dubliniensis, a new fungal pathogen
J Basic Microbiol
(2002)
H. Hof et al.
Differentiation of Candida dubliniensis from Candida albicans by means of MALDI-TOF mass spectrometry
Clin Lab
(2012)
A.M. Huang et al.
Impact of rapid organism identification via matrix-assisted laser desorption/ionization time-of-flight combined with antimicrobial stewardship team intervention in adult patients with bacteremia and candidemia
Clin Infect Dis
(2013)
P.C. Iwen et al.
Use of a panfungal PCR assay for detection of fungal pathogens in a commercial blood culture system
J Clin Microbiol
(2004)
S. Klein et al.
Integration of matrix-assisted laser desorption/ionization time-of-flight mass spectrometry in blood culture diagnostics: a fast and effective approach
J Med Microbiol
(2012)

There are more references available in the full text version of this article.

Cited by (11)

Misidentification of Candida dubliniensis isolates with the VITEK MS
2021, Journal of Medical Mycology
The phylogenetic relatedness of Candida dubliniensis and C. albicans may lead to misidentification of C. dubliniensis and underestimation of its clinical significance. We evaluated the performance of VITEK-MS in identifying C. dubliniensis isolates following growth on different culture media. Correct identification was documented in 98% of the isolates grown on blood agar media whereas only 44% were correctly identified from SDA or CHROMagar. The use of non-manufacturer validated media for identifying C. dubliniensis with VITEK-MS, may result in misidentification of these isolates as C. albicans. This finding calls for reassessing the accuracy of fungal isolates identification in local workflows using non-validated culture media.
Yeast species associated with diseased fish: Occurrence, identification, experimental challenges and antifungal susceptibility testing
2018, Aquaculture
Citation Excerpt :
Recently, several research teams have succeeded in identifying moulds using MALDI-TOF MS, which was first adapted to bacteria, then to yeast identification (Cassagne et al., 2016; Porte et al., 2017). Though there is accumulating evidence for the performance of these systems for common yeast species, there are few data for cryptic/emerging species (Roberts et al., 2016). Species identification of yeasts isolated from diseased fish using this method has seldom been reported.
The effort to characterize yeasts in fish is still meager, and identification of yeasts from aquacultured diseased fish could be a good strategy to identify those yeast species which have a role in overall fish health. Yet, knowledge on their identification using Matrix-assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF MS) is lacking. Hence, this study investigates the diversity of yeasts residing in the skin, gills, intestine and internal organs of three economically important diseased freshwater fish species. We carried out a comparative evaluation of the performance of the biochemical identification kit RapID Yeast Plus system and MALDI-TOF MS, with amplification/sequencing of Internal Transcribed Spacer (ITS1–ITS2) regions of rDNA as the reference method for yeast identification. Antifungal susceptibility pattern of yeast isolates was assessed using Vitek2 compact. Among 420 samples from Oreochromis species, Clarias gariepinus and Mugil cephalus, 215 yeast isolates (51.2%) were detected, with 7 genera: Candida, Cryptococcus, Debaryomyces, Rhodotorula, Trichosporon, Geotrichum and Pichia identified. Candida spp. accounted for 65.12% of the isolates and Rhodotorula spp. 28.84%. Overall, the yeast belonging to Ascomycota was higher than the Basidiomycota yeast (P < 0.001). There is statistically highly significant difference between incidences of yeast species in different fish. The intraperitoneal-challenged fish groups with Cryptococcus uniguttulatus, Candida lipolytica, Candida albicans and Trichosporon mucoides exhibited the highest cumulative mortality percentages (100, 80.95, 71.43 and 61.90%, respectively). C. uniguttulatus challenged group exhibited exophthalmia, opacity in the lens and yellow discoloration of the body and internal organs, meanwhile C. albicans group showed red mouth. The RapID Yeast Plus system identified the tested isolates at the species and genus level with a percentage of 30% for each. The MALDI-TOF MS accurately identified 75% of the isolates to the species level. Of the isolates (25%) not present in the MALDI-TOF MS database at the species level, 10% were ascribed to the genus level and an incorrect identification was given for 15%. Voriconazole, amphotericin B and caspofungin were the most active drugs against all the species tested, while fluconazole demonstrated poor activity. The diseased freshwater fish species may harbor potential clinically relevant yeast species, including C. albicans, non-albicans Candida species, Trichosporon, Cryptococcus and Rhodotorula species. MALDI-TOF MS offers an accurate and cost-effective alternative to the conventional and molecular methods for yeast identification. Subsequently, implementation of the VITEK MS with a more extended database is required.
Identification of cryptic Candida species by MALDI-TOF mass spectrometry, not all MALDI-TOF systems are the same: focus on the C. parapsilosis species complex
2016, Diagnostic Microbiology and Infectious Disease
Identification of Candida yeasts: Conventional methods and MALDI-TOF MS
2018, Revista Iberoamericana de Micologia
Las infecciones fúngicas invasivas han ido aumentando y las levaduras del género Candida son la principal causa de las mismas. Las especies distintas de Candida albicans son cada vez más frecuentes y algunas pueden presentar patrones variables de sensibilidad a los antimicóticos, por lo que es importante la correcta identificación de la especie. Los métodos de identificación convencionales con los que cuentan la mayoría de los laboratorios pueden presentar inconvenientes. La espectrometría de masas (MALDI-TOF MS) surgió como una alternativa a los mismos.
Evaluar la concordancia de los resultados de identificación de diferentes especies de Candida mediante dos técnicas: una convencional (galerías API) y MALDI-TOF MS.
Se analizaron las siguientes especies y número de aislamientos: Candida parapsilosis (28), Candida glabrata (34), Candida krusei (24), Candida tropicalis (45), Candida guilliermondii (30), C. albicans (28), Candida dubliniensis (6), Candida kefyr (1) y Candida lipolytica (1) del cepario de RMCABA; también se utilizaron las cepas C. parapsilosis 22019, C. glabrata 90030, C. krusei 6258 y C. albicans 68548, pertenecientes a la American Type Culture Collection (ATCC). Las discrepancias en la identificación se resolvieron mediante genotipificación.
La concordancia directa entre la identificación convencional y MALDI-TOF MS fue del 92,5% (186/201).
Invasive fungal infections are increasing, and Candida yeasts are the main cause. Species other than Candida albicans are becoming more frequent, and some of them may have variable patterns of susceptibility to antifungal agents, making it important to identify them correctly. Conventional identification methods used by most laboratories may present with drawbacks. Mass spectrometry (MALDI-TOF MS) has emerged as an alternative method.
The aim of this study was to evaluate the concordance of the identification, at species level, by conventional methods (API) and MALDI-TOF MS.
The following species and number of isolates were studied: Candida parapsilosis (28), Candida glabrata (34), Candida krusei (24), Candida tropicalis (45), Candida guilliermondii (30), C. albicans (28), Candida dubliniensis (6), Candida kefyr (1), and Candida lipolytica (1) from the strain collection of Autonomous City of Buenos Aires Mycology Network (RMCABA). The strains C. parapsilosis 22019, C. glabrata 90030, C. krusei 6258 and C. albicans 68548 from the American Type Culture Collection (ATCC) were also included. Discrepancies were resolved by genotyping.
The direct concordance between the conventional identification method and MALDI-TOF MS was 92.5% (186/201).
Mass spectrometry in research laboratories and clinical diagnostic: a new era in medical mycology
2022, Brazilian Journal of Microbiology
Candida albicans and Candida dubliniensis Show Different Trailing Effect Patterns When Exposed to Echinocandins and Azoles
2020, Frontiers in Microbiology

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Diagnostic Microbiology and Infectious Disease

Highlights

Abstract

Section snippets

Acknowledgements

References (16)

Comparative evaluation of two matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) systems for the identification of clinically significant yeasts

Int J Infect Dis

Performance of matrix-assisted laser desorption ionization-time of flight mass spectrometry for identification of bacterial strains routinely isolated in a clinical microbiology laboratory

J Clin Microbiol

Arachidonic acid increases antifungal susceptibility of Candida albicans and Candida dubliniensis

J Antimicrob Chemother

Candida dubliniensis, a new fungal pathogen

J Basic Microbiol

Differentiation of Candida dubliniensis from Candida albicans by means of MALDI-TOF mass spectrometry

Clin Lab

Impact of rapid organism identification via matrix-assisted laser desorption/ionization time-of-flight combined with antimicrobial stewardship team intervention in adult patients with bacteremia and candidemia

Clin Infect Dis

Use of a panfungal PCR assay for detection of fungal pathogens in a commercial blood culture system

J Clin Microbiol

Integration of matrix-assisted laser desorption/ionization time-of-flight mass spectrometry in blood culture diagnostics: a fast and effective approach

J Med Microbiol

Cited by (11)

Misidentification of Candida dubliniensis isolates with the VITEK MS

Yeast species associated with diseased fish: Occurrence, identification, experimental challenges and antifungal susceptibility testing

Identification of cryptic Candida species by MALDI-TOF mass spectrometry, not all MALDI-TOF systems are the same: focus on the C. parapsilosis species complex

Identification of Candida yeasts: Conventional methods and MALDI-TOF MS

Mass spectrometry in research laboratories and clinical diagnostic: a new era in medical mycology

Candida albicans and Candida dubliniensis Show Different Trailing Effect Patterns When Exposed to Echinocandins and Azoles

MycologyEvaluation of matrix-assisted laser desorption ionization–time-of-flight mass spectrometry to differentiate between Candida albicans and Candida dubliniensis

Highlights

Abstract

Section snippets

Acknowledgements

Int J Infect Dis

Performance of matrix-assisted laser desorption ionization-time of flight mass spectrometry for identification of bacterial strains routinely isolated in a clinical microbiology laboratory

J Clin Microbiol

Arachidonic acid increases antifungal susceptibility of Candida albicans and Candida dubliniensis

J Antimicrob Chemother

Candida dubliniensis, a new fungal pathogen

J Basic Microbiol

Differentiation of Candida dubliniensis from Candida albicans by means of MALDI-TOF mass spectrometry

Clin Lab

Impact of rapid organism identification via matrix-assisted laser desorption/ionization time-of-flight combined with antimicrobial stewardship team intervention in adult patients with bacteremia and candidemia

Clin Infect Dis

Use of a panfungal PCR assay for detection of fungal pathogens in a commercial blood culture system

J Clin Microbiol

Integration of matrix-assisted laser desorption/ionization time-of-flight mass spectrometry in blood culture diagnostics: a fast and effective approach

J Med Microbiol

Mycology
Evaluation of matrix-assisted laser desorption ionization–time-of-flight mass spectrometry to differentiate between Candida albicans and Candida dubliniensis