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

2017 ◽  
Vol 55 (8) ◽  
pp. 2521-2528 ◽  
Author(s):  
Mark Fraser ◽  
Andrew M. Borman ◽  
Elizabeth M. Johnson
Keyword(s):  
Mass Spectrometry ◽  
Laser Desorption ◽  
Laser Desorption Ionization ◽  
Accurate Identification ◽  
Ionization Time ◽  
Content Type ◽  
Desorption Ionization ◽  
Maldi Tof ◽  
Matrix Assisted Laser ◽  
Matrix Assisted Laser Desorption

ABSTRACTEumycetoma, 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 includeMadurellaspp.,Falciformisporaspp.,Trematosphaeria grisea,Nigrograna mackinnonii,Pseudochaetosphaeronema larense,Medicopsis romeroi, andEmarelliaspp.Rhytidhysteron rufulumandParathyridaria percutaneacause 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.

scholarly journals Performance of Matrix-Assisted Laser Desorption Ionization–Time of Flight Mass Spectrometry for Identifying Clinical Malassezia Isolates

2016 ◽  
Vol 55 (1) ◽  
pp. 90-96 ◽  
Author(s):  
Julie Denis ◽  
Marie Machouart ◽  
Florent Morio ◽  
Marcela Sabou ◽  
Catherine Kauffmann-LaCroix ◽  
...  
Keyword(s):  
Mass Spectrometry ◽  
Time Of Flight ◽  
Laser Desorption ◽  
Laser Desorption Ionization ◽  
Ionization Time ◽  
Content Type ◽  
Desorption Ionization ◽  
Maldi Tof ◽  
Matrix Assisted Laser ◽  
Matrix Assisted Laser Desorption

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.

scholarly journals Efficient Detection of Carbapenemase Activity in Enterobacteriaceae by Matrix-Assisted Laser Desorption Ionization−Time of Flight Mass Spectrometry in Less Than 30 Minutes

2015 ◽  
Vol 53 (7) ◽  
pp. 2163-2171 ◽  
Author(s):  
Camille Lasserre ◽  
Luc De Saint Martin ◽  
Gaelle Cuzon ◽  
Pierre Bogaerts ◽  
Estelle Lamar ◽  
...  
Keyword(s):  
Mass Spectrometry ◽  
Time Of Flight ◽  
Laser Desorption ◽  
Laser Desorption Ionization ◽  
Ionization Time ◽  
Content Type ◽  
Desorption Ionization ◽  
Maldi Tof ◽  
Matrix Assisted Laser ◽  
Matrix Assisted Laser Desorption

The recognition of carbapenemase-producingEnterobacteriaceae(CPE) isolates is a major laboratory challenge, and their inappropriate or delayed detection may have negative impacts on patient management and on the implementation of infection control measures. We describe here a matrix-assisted laser desorption ionization−time of flight (MALDI-TOF)-based method to detect carbapenemase activity inEnterobacteriaceae. After a 20-min incubation of the isolate with 0.5 mg/ml imipenem at 37°C, supernatants were analyzed by MALDI-TOF in order to identify peaks corresponding to imipenem (300 Da) and an imipenem metabolite (254 Da). A total of 223 strains, 77 CPE (OXA-48 variants, KPC, NDM, VIM, IMI, IMP, and NMC-A) and 146 non-CPE (cephalosporinases, extended-spectrum β-lactamases [ESBLs], and porin defects), were tested and used to calculate a ratio of imipenem hydrolysis: mass spectrometry [MS] ratio = metabolite/(imipenem + metabolite). An MS ratio cutoff was statistically determined to classify strains as carbapenemase producers (MS ratio of ≥0.82). We validated this method first by testing 30 of our 223 isolates (15 CPE and 15 non-CPE) 10 times to calculate an intraclass correlation coefficient (ICC of 0.98), showing the excellent repeatability of the method. Second, 43 strains (25 CPE and 18 non-CPE) different from the 223 strains used to calculate the ratio cutoff were used as external controls and blind tested. They yielded sensitivity and specificity of 100%. The total cost per test is <0.10 U.S. dollars (USD). This easy-to-perform assay is time-saving, cost-efficient, and highly reliable and might be used in any routine laboratory, given the availability of mass spectrometry, to detect CPE.

scholarly journals Creation of an In-House Matrix-Assisted Laser Desorption Ionization–Time of Flight Mass Spectrometry Corynebacterineae Database Overcomes Difficulties in Identification of Nocardia farcinica Clinical Isolates

2015 ◽  
Vol 53 (8) ◽  
pp. 2611-2621 ◽  
Author(s):  
Mariola Paściak ◽  
Władysław Dacko ◽  
Joanna Sikora ◽  
Danuta Gurlaga ◽  
Krzysztof Pawlik ◽  
...  
Keyword(s):  
Mass Spectrometry ◽  
Time Of Flight ◽  
Laser Desorption ◽  
Laser Desorption Ionization ◽  
Ionization Time ◽  
Content Type ◽  
Desorption Ionization ◽  
Maldi Tof ◽  
Matrix Assisted Laser ◽  
Matrix Assisted Laser Desorption

Nocardiosis is a rare disease that is caused by Gram-positive actinobacteria of theNocardiagenus and affects predominantly immunocompromised patients. In its disseminated form, it has a predilection for the central nervous system and is associated with high mortality rates. Therefore, prompt identification of the pathogen is critical. Matrix-assisted laser desorption ionization–time of flight (MALDI-TOF) mass spectrometry is a relatively novel technique used for identification of microorganisms. In this work, an upgraded MALDI-TOF Biotyper database containingCorynebacterineaerepresentatives of strains deposited in the Polish Collection of Microorganisms was created and used for identification of the strain isolated from a nocardial brain abscess, mimicking a brain tumor, in an immunocompetent patient. Testing with the API Coryne system initially incorrectly identifiedRhodococcussp., while chemotaxonomic tests, especially mycolic acid analysis, enabled correctNocardiaidentification only at the genus level. Subsequent sequence analysis of 16S rRNA andsecA1genes confirmed the identification. To improve the accuracy of the results, an in-house database was constructed using optimized parameters; with the use of the database, the strain was eventually identified asNocardia farcinica. Clinical laboratories processing various clinical strains can upgrade a commercial database to improve and to accelerate the results obtained. This is especially important in the case ofNocardia, for which valid microbial diagnosis remains challenging; reference laboratories are often required to identify and to survey these rare actinobacteria.

scholarly journals Misidentification of a Rare Species, Cryptococcus laurentii, by Commonly Used Commercial Biochemical Methods and Matrix-Assisted Laser Desorption Ionization–Time of Flight Mass Spectrometry Systems: Challenges for Clinical Mycology Laboratories

2015 ◽  
Vol 54 (1) ◽  
pp. 226-229 ◽  
Author(s):  
Meng Xiao ◽  
Xin Fan ◽  
Xin-Xin Chen ◽  
He Wang ◽  
Li Zhang ◽  
...  
Keyword(s):  
Mass Spectrometry ◽  
Laser Desorption ◽  
Cryptococcus Laurentii ◽  
Laser Desorption Ionization ◽  
Ionization Time ◽  
Content Type ◽  
Desorption Ionization ◽  
Flight Mass Spectrometry ◽  
Matrix Assisted Laser ◽  
Matrix Assisted Laser Desorption

Forty-two putativeCryptococcus laurentiiisolates identified by the Vitek 2 system were collected in China. The gold standard, internal transcribed spacer (ITS) sequencing, confirmed that only two isolates were genuineC. laurentii. Bruker Biotyper matrix-assisted laser desorption ionization–time of flight mass spectrometry was able to identify theC. laurentiiisolates with an expanded custom database.

scholarly journals Beyond the Matrix-Assisted Laser Desorption Ionization (MALDI) Biotyping Workflow: in Search of Microorganism-Specific Tryptic Peptides Enabling Discrimination of Subspecies

2014 ◽  
Vol 80 (14) ◽  
pp. 4234-4241 ◽  
Author(s):  
Maria-Theresia Gekenidis ◽  
Patrick Studer ◽  
Simone Wüthrich ◽  
René Brunisholz ◽  
David Drissner
Keyword(s):  
Mass Spectrometry ◽  
Laser Desorption ◽  
Clinical Diagnostics ◽  
Laser Desorption Ionization ◽  
Accurate Identification ◽  
Discrimination Power ◽  
Desorption Ionization ◽  
Maldi Tof ◽  
Matrix Assisted Laser ◽  
Matrix Assisted Laser Desorption

ABSTRACTA well-accepted method for identification of microorganisms uses matrix-assisted laser desorption ionization–time of flight mass spectrometry (MALDI-TOF MS) coupled to analysis software which identifies and classifies the organism according to its ribosomal protein spectral profile. The method, called MALDI biotyping, is widely used in clinical diagnostics and has partly replaced conventional microbiological techniques such as biochemical identification due to its shorter time to result (minutes for MALDI biotyping versus hours or days for classical phenotypic or genotypic identification). Besides its utility for identifying bacteria, MS-based identification has been shown to be applicable also to yeasts and molds. A limitation to this method, however, is that accurate identification is most reliably achieved on the species level on the basis of reference mass spectra, making further phylogenetic classification unreliable. Here, it is shown that combining tryptic digestion of the acid/organic solvent extracted (classical biotyping preparation) and resolubilized proteins, nano-liquid chromatography (nano-LC), and subsequent identification of the peptides by MALDI-tandem TOF (MALDI-TOF/TOF) mass spectrometry increases the discrimination power to the level of subspecies. As a proof of concept, using this targeted proteomics workflow, we have identified subspecies-specific biomarker peptides for threeSalmonellasubspecies, resulting in an extension of the mass range and type of proteins investigated compared to classical MALDI biotyping. This method therefore offers rapid and cost-effective identification and classification of microorganisms at a deeper taxonomic level.

scholarly journals Identification of Neisseria gonorrhoeae by the Bruker Biotyper Matrix-Assisted Laser Desorption Ionization–Time of Flight Mass Spectrometry System Is Improved by a Database Extension: TABLE 1

2016 ◽  
Vol 54 (4) ◽  
pp. 1130-1132 ◽  
Author(s):  
Valentijn A. Schweitzer ◽  
Alje P. van Dam ◽  
I Putu Yuda Hananta ◽  
Rob Schuurman ◽  
Johannes G. Kusters ◽  
...  
Keyword(s):  
Mass Spectrometry ◽  
Neisseria Gonorrhoeae ◽  
Laser Desorption ◽  
Laser Desorption Ionization ◽  
Ionization Time ◽  
Content Type ◽  
Desorption Ionization ◽  
Flight Mass Spectrometry ◽  
Matrix Assisted Laser ◽  
Matrix Assisted Laser Desorption

Identification ofNeisseria gonorrhoeaeby the Bruker matrix-assisted laser desorption ionization−time of flight mass spectrometry (MALDI-TOF MS) system may be affected by “B consistency categorization.” A supplementary database of 17N. gonorrhoeaemain spectra was constructed. Twelve of 64N. gonorrhoeaeidentifications were categorized with B consistency, which disappeared using the supplementary database. Database extension did not result in misidentification ofNeisseria meningitidis.

scholarly journals A Comprehensive Evaluation of the Bruker Biotyper MS and Vitek MS Matrix-Assisted Laser Desorption Ionization–Time of Flight Mass Spectrometry Systems for Identification of Yeasts, Part of the National China Hospital Invasive Fungal Surveillance Net (CHIF-NET) Study, 2012 to 2013

2016 ◽  
Vol 54 (5) ◽  
pp. 1376-1380 ◽  
Author(s):  
He Wang ◽  
Yan-Yan Fan ◽  
Timothy Kudinha ◽  
Zhi-Peng Xu ◽  
Meng Xiao ◽  
...  
Keyword(s):  
Mass Spectrometry ◽  
Laser Desorption ◽  
Laser Desorption Ionization ◽  
Ionization Time ◽  
Content Type ◽  
Desorption Ionization ◽  
Flight Mass Spectrometry ◽  
Vitek Ms ◽  
Matrix Assisted Laser ◽  
Matrix Assisted Laser Desorption

Among the 2,683 yeast isolates representing 41 different species (25CandidaandCandida-related species and 16 non-Candidayeast species) collected in the National China Hospital Invasive Fungal Surveillance Net (CHIF-NET) program (2012 to 2013), the Bruker Biotyper MS matrix-assisted laser desorption ionization–time of flight mass spectrometry (MALDI-TOF MS) system exhibited significantly higher accuracy rates than the Vitek MS system for identification of all yeast isolates (98.8% versus 95.4%,P<0.001 by Pearson's chi-square test) and for allCandidaandCandida-related species isolates (99.4% versus 95.5%,P< 0.001).

Can Matrix-Assisted Laser Desorption Ionization Time-of-Flight Mass Spectrometry (MALDI-TOF) Enhance Antimicrobial Stewardship Efforts in the Acute Care Setting?

2013 ◽  
Vol 34 (9) ◽  
pp. 990-995 ◽  
Author(s):  
Pranita D. Tamma ◽  
Kennard Tan ◽  
Veronique R. Nussenblatt ◽  
Alison E. Turnbull ◽  
Karen C. Carroll ◽  
...  
Keyword(s):  
Mass Spectrometry ◽  
Time Of Flight ◽  
Laser Desorption ◽  
Laser Desorption Ionization ◽  
Ionization Time ◽  
Desorption Ionization ◽  
Maldi Tof ◽  
Flight Mass Spectrometry ◽  
Matrix Assisted Laser ◽  
Matrix Assisted Laser Desorption

We evaluated 222 hospitalized patients whose clinical isolates were tested using standard methods and matrix-assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-TOF). MALDI-TOF could have reduced time to appropriate therapy for 28.8% and 44.6% patients based on the treating physician's choices and stewardship team recommendations, respectively. Clinicians should be aware of scenarios in which MALDI-TOF can optimize antibiotic therapy.

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