scholarly journals Performance of Matrix-Assisted Laser Desorption Ionization−Time of Flight Mass Spectrometry for Identification of Aspergillus, Scedosporium, and Fusarium spp. in the Australian Clinical Setting

2016 ◽  
Vol 54 (8) ◽  
pp. 2182-2186 ◽  
Author(s):  
Sue Sleiman ◽  
Catriona L. Halliday ◽  
Belinda Chapman ◽  
Mitchell Brown ◽  
Joanne Nitschke ◽  
...  

We developed an Australian database for the identification ofAspergillus,Scedosporium, andFusariumspecies (n= 28) by matrix-assisted laser desorption ionization−time of flight mass spectrometry (MALDI-TOF MS). In a challenge against 117 isolates, species identification significantly improved when the in-house-built database was combined with the Bruker Filamentous Fungi Library compared with that for the Bruker library alone (Aspergillus, 93% versus 69%;Fusarium, 84% versus 42%; andScedosporium, 94% versus 18%, respectively).

2016 ◽  
Vol 54 (9) ◽  
pp. 2354-2364 ◽  
Author(s):  
Aradhana Masih ◽  
Pradeep K. Singh ◽  
Shallu Kathuria ◽  
Kshitij Agarwal ◽  
Jacques F. Meis ◽  
...  

Aspergillusspecies cause a wide spectrum of clinical infections. AlthoughAspergillus fumigatusandAspergillus flavusremain the most commonly isolated species in aspergillosis, in the last decade, rare and crypticAspergillusspecies have emerged in diverse clinical settings. The present study analyzed the distribution andin vitroantifungal susceptibility profiles of rareAspergillusspecies in clinical samples from patients with suspected aspergillosis in 8 medical centers in India. Further, a matrix-assisted laser desorption ionization–time of flight mass spectrometry in-house database was developed to identify these clinically relevantAspergillusspecies. β-Tubulin and calmodulin gene sequencing identified 45 rareAspergillusisolates to the species level, except for a solitary isolate. They included 23 less commonAspergillusspecies belonging to 12 sections, mainly inCircumdati,Nidulantes,Flavi,Terrei,Versicolores,Aspergillus, andNigri. Matrix-assisted laser desorption ionization–time of flight mass spectrometry (MALDI-TOF MS) identified only 8 (38%) of the 23 rareAspergillusisolates to the species level. Following the creation of an in-house database with the remaining 14 species not available in the Bruker database, the MALDI-TOF MS identification rate increased to 95%. Overall, high MICs of ≥2 μg/ml were noted for amphotericin B in 29% of the rareAspergillusspecies, followed by voriconazole in 20% and isavuconazole in 7%, whereas MICs of >0.5 μg/ml for posaconazole were observed in 15% of the isolates. Regarding the clinical diagnoses in 45 patients with positive rareAspergillusspecies cultures, 19 (42%) were regarded to represent colonization. In the remaining 26 patients, rareAspergillusspecies were the etiologic agent of invasive, chronic, and allergic bronchopulmonary aspergillosis, allergic fungal rhinosinusitis, keratitis, and mycetoma.


2000 ◽  
Vol 66 (11) ◽  
pp. 4720-4724 ◽  
Author(s):  
Matthew L. Magnuson ◽  
James H. Owens ◽  
Catherine A. Kelty

ABSTRACT Matrix-assisted laser desorption ionization–time of flight mass spectrometry (MALDI-TOF MS) was used to investigate whole and freeze-thawed Cryptosporidium parvum oocysts. Whole oocysts revealed some mass spectral features. Reproducible patterns of spectral markers and increased sensitivity were obtained after the oocysts were lysed with a freeze-thaw procedure. Spectral-marker patterns forC. parvum were distinguishable from those obtained forCryptosporidium muris. One spectral marker appears specific for the genus, while others appear specific at the species level. Three different C. parvum lots were investigated, and similar spectral markers were observed in each. Disinfection of the oocysts reduced and/or eliminated the patterns of spectral markers.


2016 ◽  
Vol 55 (1) ◽  
pp. 90-96 ◽  
Author(s):  
Julie Denis ◽  
Marie Machouart ◽  
Florent Morio ◽  
Marcela Sabou ◽  
Catherine Kauffmann-LaCroix ◽  
...  

ABSTRACT The genus Malassezia comprises commensal yeasts on human skin. These yeasts are involved in superficial infections but are also isolated in deeper infections, such as fungemia, particularly in certain at-risk patients, such as neonates or patients with parenteral nutrition catheters. Very little is known about Malassezia epidemiology and virulence. This is due mainly to the difficulty of distinguishing species. Currently, species identification is based on morphological and biochemical characteristics. Only molecular biology techniques identify species with certainty, but they are time-consuming and expensive. The aim of this study was to develop and evaluate a matrix-assisted laser desorption ionization–time of flight (MALDI-TOF) database for identifying Malassezia species by mass spectrometry. Eighty-five Malassezia isolates from patients in three French university hospitals were investigated. Each strain was identified by internal transcribed spacer sequencing. Forty-five strains of the six species Malassezia furfur , M. sympodialis , M. slooffiae , M. globosa , M. restricta , and M. pachydermatis allowed the creation of a MALDI-TOF database. Forty other strains were used to test this database. All strains were identified by our Malassezia database with log scores of >2.0, according to the manufacturer's criteria. Repeatability and reproducibility tests showed a coefficient of variation of the log score values of <10%. In conclusion, our new Malassezia database allows easy, fast, and reliable identification of Malassezia species. Implementation of this database will contribute to a better, more rapid identification of Malassezia species and will be helpful in gaining a better understanding of their epidemiology.


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