Identification of Candida albicans species complex in vulvovaginal candidiasis by using conventional methods, hwp1 gene polymorphisms and MALDI-TOF MS profiles

2018 ◽  
Author(s):  
E. Nasri
Keyword(s):  
Candida Albicans ◽  
Gene Polymorphisms ◽  
Species Complex ◽  
Vulvovaginal Candidiasis ◽  
Maldi Tof Ms ◽  
Maldi Tof ◽  
Conventional Methods ◽  
Tof Ms

scholarly journals Testing the Applicability of MALDI-TOF MS as an Alternative Stock Identification Method in a Cryptic Species Complex

Molecules ◽  
2020 ◽  
Vol 25 (14) ◽  
pp. 3214
Author(s):  
Gabor Maasz ◽  
Zita Zrínyi ◽  
Istvan Fodor ◽  
Nóra Boross ◽  
Zoltán Vitál ◽  
...  
Keyword(s):  
Sample Preparation ◽  
Cryptic Species ◽  
Species Complex ◽  
Intraspecific Variability ◽  
Clove Oil ◽  
Term Survival ◽  
Maldi Tof Ms ◽  
Maldi Tof ◽  
Ms Analysis ◽  
Tof Ms

Knowledge of intraspecific variability of a certain species is essential for their long-term survival and for the development of conservation plans. Nowadays, molecular/genetic methods are the most frequently used for this purpose. Although, the Matrix Assisted Laser Desorption Ionization Time of Flight Mass Spectrometry (MALDI-TOF MS) technique has become a promising alternative tool to specify intraspecific variability, there is a lack of information about the limitations of this method, and some methodological issues need to be resolved. Towards this goal, we tested the sensitivity of this method on an intraspecific level, using genetically identified individuals of a cryptic fish species complex collected from five distinct populations. Additionally, some methodologic issues, such as the effect of (1) delayed sample preparation, (2) clove oil anaesthetization, and (3) different tissue types (muscle, and brain) were investigated using the MS analysis results. Our results show that the delayed sample preparation has a fundamental effect on the result of MS analysis, while at the same time the clove oil did not affect the results considerably. Both the brain and muscle samples were usable for cryptic species identification, but in our opinion this method has limited applicability for population-level segregation. The application of MALDI-TOF MS to the exploitable toolkit of phylogenetic and taxonomic researches could be used to broaden conclusions.

scholarly journals Evaluation of the new Id‐Fungi plates from Conidia for MALDI‐TOF MS identification of filamentous fungi and comparison with conventional methods as identification tool for dermatophytes from nails, hair and skin samples

Mycoses ◽  
2020 ◽  
Vol 63 (10) ◽  
pp. 1115-1127 ◽  
Author(s):  
Rosalie Sacheli ◽  
Anne‐Sophie Henri ◽  
Laurence Seidel ◽  
Marie Ernst ◽  
Rajae Darfouf ◽  
...  
Keyword(s):  
Filamentous Fungi ◽  
Maldi Tof Ms ◽  
Maldi Tof ◽  
Conventional Methods ◽  
Skin Samples ◽  
Tof Ms ◽  
Identification Tool

scholarly journals Determination of Drug Efflux Pump Efficiency in Drug-Resistant Bacteria Using MALDI-TOF MS

Antibiotics ◽  
2020 ◽  
Vol 9 (10) ◽  
pp. 639 ◽  
Author(s):  
Wen-Jung Lu ◽  
Hsuan-Ju Lin ◽  
Pang-Hung Hsu ◽  
Hong-Ting Victor Lin
Keyword(s):  
Minimum Inhibitory Concentration ◽  
Inhibitory Concentration ◽  
Resistant Bacteria ◽  
Pump Efficiency ◽  
Drug Efflux ◽  
Maldi Tof Ms ◽  
E Coli ◽  
Maldi Tof ◽  
Conventional Methods ◽  
Tof Ms

Multidrug efflux pumps play an essential role in antibiotic resistance. The conventional methods, including minimum inhibitory concentration and fluorescent assays, to monitor transporter efflux activity might have some drawbacks, such as indirect evidence or interference from color molecules. In this study, MALDI-TOF MS use was explored for monitoring drug efflux by a multidrug transporter, and the results were compared for validation with the data from conventional methods. Minimum inhibitory concentration was used first to evaluate the activity of Escherichia coli drug transporter AcrB, and this analysis showed that the E. coli overexpressing AcrB exhibited elevated resistance to various antibiotics and dyes. Fluorescence-based studies indicated that AcrB in E. coli could decrease the accumulation of intracellular dyes and display various efflux rate constants for different dyes, suggesting AcrB’s efflux activity. The MALDI-TOF MS analysis parameters were optimized to maintain a detection accuracy for AcrB’s substrates; furthermore, the MS data showed that E. coli overexpressing AcrB led to increased ions abundancy of various dyes and drugs in the extracellular space at different rates over time, illustrating continuous substrate efflux by AcrB. This study concluded that MALDI-TOF MS is a reliable method that can rapidly determine the drug pump efflux activity for various substrates.

scholarly journals Mass Spectrometry Proteotyping-Based Detection and Identification of Staphylococcus aureus, Escherichia coli, and Candida albicans in Blood

2021 ◽  
Vol 11 ◽  
Author(s):  
Nahid Kondori ◽  
Amra Kurtovic ◽  
Beatriz Piñeiro-Iglesias ◽  
Francisco Salvà-Serra ◽  
Daniel Jaén-Luchoro ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Staphylococcus Aureus ◽  
Candida Albicans ◽  
Blood Cultures ◽  
Blood Samples ◽  
Maldi Tof Ms ◽  
E Coli ◽  
Maldi Tof ◽  
Detection And Identification ◽  
Tof Ms

Bloodstream infections (BSIs), the presence of microorganisms in blood, are potentially serious conditions that can quickly develop into sepsis and life-threatening situations. When assessing proper treatment, rapid diagnosis is the key; besides clinical judgement performed by attending physicians, supporting microbiological tests typically are performed, often requiring microbial isolation and culturing steps, which increases the time required for confirming positive cases of BSI. The additional waiting time forces physicians to prescribe broad-spectrum antibiotics and empirically based treatments, before determining the precise cause of the disease. Thus, alternative and more rapid cultivation-independent methods are needed to improve clinical diagnostics, supporting prompt and accurate treatment and reducing the development of antibiotic resistance. In this study, a culture-independent workflow for pathogen detection and identification in blood samples was developed, using peptide biomarkers and applying bottom-up proteomics analyses, i.e., so-called “proteotyping”. To demonstrate the feasibility of detection of blood infectious pathogens, using proteotyping, Escherichia coli and Staphylococcus aureus were included in the study, as the most prominent bacterial causes of bacteremia and sepsis, as well as Candida albicans, one of the most prominent causes of fungemia. Model systems including spiked negative blood samples, as well as positive blood cultures, without further culturing steps, were investigated. Furthermore, an experiment designed to determine the incubation time needed for correct identification of the infectious pathogens in blood cultures was performed. The results for the spiked negative blood samples showed that proteotyping was 100- to 1,000-fold more sensitive, in comparison with the MALDI-TOF MS-based approach. Furthermore, in the analyses of ten positive blood cultures each of E. coli and S. aureus, both the MALDI-TOF MS-based and proteotyping approaches were successful in the identification of E. coli, although only proteotyping could identify S. aureus correctly in all samples. Compared with the MALDI-TOF MS-based approaches, shotgun proteotyping demonstrated higher sensitivity and accuracy, and required significantly shorter incubation time before detection and identification of the correct pathogen could be accomplished.

scholarly journals Use of Matrix-Assisted Laser Desorption Ionization–Time of Flight Mass Spectrometry for Identification of Molds of the Fusarium Genus

2014 ◽  
Vol 53 (2) ◽  
pp. 465-476 ◽  
Author(s):  
David Triest ◽  
Dirk Stubbe ◽  
Koen De Cremer ◽  
Denis Piérard ◽  
Anne-Cécile Normand ◽  
...  
Keyword(s):  
Species Complex ◽  
Laser Desorption ◽  
Laser Desorption Ionization ◽  
Ionization Time ◽  
Content Type ◽  
Maldi Tof Ms ◽  
Maldi Tof ◽  
Flight Mass Spectrometry ◽  
Species Complexes ◽  
Tof Ms

The rates of infection withFusariummolds are increasing, and a diverse number ofFusariumspp. belonging to different species complexes can cause infection. Conventional species identification in the clinical laboratory is time-consuming and prone to errors. We therefore evaluated whether matrix-assisted laser desorption ionization–time of flight mass spectrometry (MALDI-TOF MS) is a useful alternative. The 289Fusariumstrains from the Belgian Coordinated Collections of Microorganisms (BCCM)/Institute of Hygiene and Epidemiology Mycology (IHEM) culture collection with validated sequence-based identities and comprising 40 species were used in this study. An identification strategy was developed, applying a standardized MALDI-TOF MS assay and an in-house reference spectrum database.In vitroantifungal testing was performed to assess important differences in susceptibility between clinically relevant species/species complexes. We observed that no incorrect species complex identifications were made by MALDI-TOF MS, and 82.8% of the identifications were correct to the species level. This success rate was increased to 91% by lowering the cutoff for identification. Although the identification of the correct species complex member was not always guaranteed, antifungal susceptibility testing showed that discriminating betweenFusariumspecies complexes can be important for treatment but is not necessarily required between members of a species complex. With this perspective, someFusariumspecies complexes with closely related members can be considered as a whole, increasing the success rate of correct identifications to 97%. The application of our user-friendly MALDI-TOF MS identification approach resulted in a dramatic improvement in both time and accuracy compared to identification with the conventional method. A proof of principle of our MALDI-TOF MS approach in the clinical setting using recently isolatedFusariumstrains demonstrated its validity.

scholarly journals MALDI-TOF MS: the proteomic approach, the future of fungal identification in India

2020 ◽  
Vol 8 (7) ◽  
pp. 2575
Author(s):  
Mallika Fonseca ◽  
Sarala Menon ◽  
Praveen Rahi ◽  
Prachi Patekar ◽  
Abhay S. Chowdhary
Keyword(s):  
Yeast Species ◽  
Fungal Infections ◽  
Tertiary Care ◽  
Turnaround Time ◽  
Maldi Tof Ms ◽  
Maldi Tof ◽  
Flight Mass Spectrometry ◽  
Proteomic Approach ◽  
Conventional Methods ◽  
Tof Ms

Background: India, being a country where fungal infections are rampant, is urgently in need of effective tools for early and accurate diagnosis of fungal infections. Matrix-assisted Laser Desorption Ionization Time-of-Flight Mass Spectrometry (MALDI-TOF-MS) is a recent method which has shown potential in identifying clinically important bacterial pathogens as well as clinically important fungi. The main objective of this study was to compare the utility of MALDI-TOF MS for the identification of fungi against that of conventional methods.Methods: The project was carried out in a tertiary care government hospital in India. Fifty clinical isolates comprising mainly various yeast species were subjected to conventional identification (Phenotypic) as well as MALDI-TOF-MS. Their results were further compared.Results: MALDI-TOF MS showed a high concordance with conventional methods while identifying species like C. albicans, C. tropicalis, C. parapsilosis and C. neoformans, although the concordance for species such as Rhodotorula and Trichosporon could only be matched up to genus level.Conclusions: MALDI-TOF MS-based identification is both a rapid and a viable tool for identification of clinically relevant yeast species with good correlation to conventional methods and a quick turnaround time.

scholarly journals Identification of Candida parapsilosis Species Complex and Lodderomyces elongisporus Isolates with MALDI-TOF MS

2021 ◽  
Author(s):  
Engin Kaplan ◽  
Ayşe Sultan Karakoyun ◽  
Deniz Alkaya ◽  
Nevzat Ünal ◽  
Aylin Döğen ◽  
...  
Keyword(s):  
Species Complex ◽  
Candida Parapsilosis ◽  
Antifungal Susceptibility ◽  
Maldi Tof Ms ◽  
Biochemical Methods ◽  
Maldi Tof ◽  
Candida Orthopsilosis ◽  
Susceptibility Profiles ◽  
Reference Strains ◽  
Tof Ms

Objective: Candida parapsilosis species complex and Lodderomyces elongisporus may have differences in terms of their virulence, prevalence, and antifungal susceptibility profiles. These species are difficult to identify with biochemical methods. Therefore, there is a need for more efficient identification methods in terms of time, cost, and applicability. This study aims to evaluate the diagnostic performance of the MALDI-TOF MS method in discriminating between isolates belonging to the C. parapsilosis species complex and L. elongisporus. Method: In the current study, a total of 32 reference strains, including the C. parapsilosis (n=8), Candida orthopsilosis (n=7), Candida metapsilosis (n=6), and L. elongisporus (n=11) species were identified using the MALDI-TOF MS method. Results: The species names of 31 (93.7%) isolates belonging to the C. parapsilosis species complex and L.elongisporus were correctly identified. Twenty four isolates including eight (100%) C. parapsilosis, five (83%) C. metapsilosis, five (71%) C. orthopsilosis, and six (54%) L. elongisporus isolates were identified with score values ranging from 1.7 to 2.14. According to the secure identification reference score of ≥ 1.7, the sensitivity and specificity of the MALDI-TOF MS method were determined as 54.5–100% and 96.3–100%, respectively. Conclusion: Although the MALDI-TOF MS method has been shown to be effective in the rapid molecular phenotypic diagnosis of species that were difficult to discriminate using biochemical methods such as C. parapsilosis species complex and L. elongisporus, there is a clear need to optimize the method and develop a larger MS library for species-level identification within secure score ranges.

scholarly journals First Food-Borne Cases of Vibrio Parahaemolyticus in Turkey

ANKEM Dergisi ◽  
2021 ◽  
Author(s):  
Nilgün Kansak ◽  
Rıza Adaleti ◽  
Belkıs Levent ◽  
Sebahat Aksaray
Keyword(s):  
Vibrio Parahaemolyticus ◽  
Reference Laboratory ◽  
Seafood Consumption ◽  
Maldi Tof Ms ◽  
Maldi Tof ◽  
Food Borne ◽  
Stool Samples ◽  
Vitek 2 ◽  
Conventional Methods ◽  
Tof Ms

ABSTRACT Vibrio parahaemolyticus (V.parahaemolyticus) is detected in many parts of the world. It is one of the most common causes of food-borne infections in Asian countries and Japan, and is usually seen as minor outbreaks involving less than ten cases. In this study, it is aimed to investigate V.parahaemolyticus in diarrhea cases in our laboratory in order to draw attention to the possible cases of this agent due to the increase in the consumption of shellfish. In the period of July-August 2018; patients who applied to the emergency service of our hospital with gastrointestinal tract infection symptoms following seafood consumption were investigated by stool microscopy and culture as part of routine procedures. Stool samples were cultured on Hektoen enteric agar, MacConkey agar, and sheep blood agar and were incubated at 37°C for 24 hours. After the incubation period, lactose negative and oxidase-positive colonies were identified by classical biochemical tests, VITEK 2 and MALDI-TOF MS (bioMérieux, France). In seven patients aged between 12-59, clinical symptoms associated with gastroenteritidis started after consuming stuffed mussels in four, eating fish in one, and in a patient after consuming fast food. One patient could not be contacted. In the microscopic examination of the macroscopically watery and mucous stool samples, abundant leukocytes in all samples, and abundant erythrocytes in addition to leukocytes in one sample were seen. The bacteria grown in culture were identified as V.parahaemolyticus by conventional methods and automated systems, Vitek 2 with 96 % and MALDITOF MS with 99 % accuracy. The results were also confirmed by the General Directorate of Public Health, National Enteric Pathogens Reference Laboratory by conventional methods, API 20 E and MALDI-TOF MS (BrukerDaltonics, USA). It should be kept in mind that V.parahaemolyticus can be isolated as a cause of gastroenteritis in diarrhea cases during the summer months, especially in the presence of a history of seafood consumption, and further investigations should be performed in this direction.

MALDI-TOF MS fingerprinting for identification and differentiation of species within the Fusarium fujikuroi species complex

2019 ◽  
Vol 103 (13) ◽  
pp. 5323-5337 ◽  
Author(s):  
Évelin F. Wigmann ◽  
Jürgen Behr ◽  
Rudi F. Vogel ◽  
Ludwig Niessen
Keyword(s):  
Species Complex ◽  
Fusarium Fujikuroi ◽  
Maldi Tof Ms ◽  
Maldi Tof ◽  
Fusarium Fujikuroi Species Complex ◽  
Tof Ms

scholarly journals Multicenter Matrix-Assisted Laser Desorption Ionization–Time of Flight Mass Spectrometry Study for Identification of Clinically Relevant Nocardia spp.

2016 ◽  
Vol 54 (5) ◽  
pp. 1251-1258 ◽  
Author(s):  
Sara J. Blosser ◽  
Steven K. Drake ◽  
Jennifer L. Andrasko ◽  
Christina M. Henderson ◽  
Kamal Kamboj ◽  
...  
Keyword(s):  
Species Complex ◽  
Laser Desorption ◽  
Laser Desorption Ionization ◽  
Ionization Time ◽  
Content Type ◽  
Maldi Tof Ms ◽  
Maldi Tof ◽  
Flight Mass Spectrometry ◽  
Level Identification ◽  
Tof Ms

This multicenter study analyzedNocardiaspp., including extraction, spectral acquisition, Bruker matrix-assisted laser desorption ionization–time of flight mass spectrometry (MALDI-TOF MS) identification, and score interpretation, using threeNocardialibraries, the Bruker, National Institutes of Health (NIH), and The Ohio State University (OSU) libraries, and compared the results obtained by each center. A standardized study protocol, 150Nocardiaisolates, and NIH and OSUNocardiaMALDI-TOF MS libraries were distributed to three centers. Following standardized culture, extraction, and MALDI-TOF MS analysis, isolates were identified using score cutoffs of ≥2.0 for species/species complex-level identification and ≥1.8 for genus-level identification. Isolates yielding a score of <2.0 underwent a single repeat extraction and analysis. The overall score range for all centers was 1.3 to 2.7 (average, 2.2 ± 0.3), with common species generally producing higher average scores than less common ones. Score categorization and isolate identification demonstrated 86% agreement between centers; 118 of 150 isolates were correctly identified to the species/species complex level by all centers. Nine strains (6.0%) were not identified by any center, and six (4.0%) of these were uncommon species with limited library representation. A categorical score discrepancy among centers occurred for 21 isolates (14.0%). There was an overall benefit of 21.2% from repeat extraction of low-scoring isolates and a center-dependent benefit for duplicate spotting (range, 2 to 8.7%). Finally, supplementation of the BrukerNocardiaMALDI-TOF MS library with both the OSU and NIH libraries increased the genus-level and species-level identification by 18.2% and 36.9%, respectively. Overall, this study demonstrates the ability of diverse clinical microbiology laboratories to utilize MALDI-TOF MS for the rapid identification of clinically relevantNocardiaspp. and to implement MALDI-TOF MS libraries developed by single laboratories across institutions.

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