Identification of hypocrealean reptile pathogenic isolates with MALDI-TOF MS

Abstract Biotyper analysis of Nannizziopsis guarroi, a fatal fungal pathogen in lizards, was described recently. Hypocrealean fungal infections in captive reptiles appear with an increasing frequency during the last decade. Therefore, the aim of this study was to proof Matrix-assisted laser desorption ionization-time-of-flight mass spectrometry (MALDI-TOF MS) as diagnostic tool for the identification of reptile pathogenic hypocrealean fungi. Ten fungal isolates obtained from nine reptiles with fungal glossitis, disseminated visceral mycosis, pneumomycosis, and fungal keratitis were analyzed. Phylogeny consisted of fragments of the large subunit of nuclear encoded ribosomal DNA (D1/D2, LSU) and the internal transcribed spacer region 1 of nuclear encoded ribosomal DNA (ITS1) as well as the protein coding gene translation elongation factor 1 alpha (TEF). Results revealed unanimously two Metarhizium granulomatis genotypes in a total of three isolates, various M. viride genotypes (n = 3), two different Purpureocillium lilacinum isolates as well as one isolate of each P. lavendulum and Beauveria bassiana. Purpureocillium lilacinum and B. bassiana are likewise frequently employed as a mycoinsecticide and mycoacaricide in agriculture on a worldwide scale and have occasionally been reported in man, causing fungal keratitis, sclerokeratitis, nosocomial infections in immunosuppressed patients, as well as cavitary pulmonary disease and cutaneous hyalohyphomycosis in immunocompetent patients. According to the results establishment of Biotyper analysis for faster differentiation of reptile-associated fungal pathogens is entirely justified.

Download Full-text

Update on the Evolving Role of MALDI-TOF MS for Laboratory Diagnosis of Fungal Infections

Current Fungal Infection Reports ◽

10.1007/s12281-012-0093-y ◽

2012 ◽

Vol 6 (3) ◽

pp. 206-214 ◽

Cited By ~ 6

Author(s):

Edith Vermeulen ◽

Jan Verhaegen ◽

Christophe Indevuyst ◽

Katrien Lagrou

Keyword(s):

Fungal Infections ◽

Laboratory Diagnosis ◽

Maldi Tof Ms ◽

Maldi Tof ◽

Tof Ms

Download Full-text

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

International Journal of Research in Medical Sciences ◽

10.18203/2320-6012.ijrms20202898 ◽

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.

Download Full-text

Gongronella eborensis sp. nov., from vineyard soil of Alentejo (Portugal)

INTERNATIONAL JOURNAL OF SYSTEMATIC AND EVOLUTIONARY MICROBIOLOGY ◽

10.1099/ijsem.0.004201 ◽

2020 ◽

Vol 70 (5) ◽

pp. 3475-3482 ◽

Cited By ~ 1

Author(s):

Maria Rosário Martins ◽

Carla Santos ◽

Célia Soares ◽

Cledir Santos ◽

Nelson Lima

Keyword(s):

Ribosomal Dna ◽

Sequence Data ◽

Novel Species ◽

Molecular Data ◽

Fungal Species ◽

Proteomic Profiling ◽

Maldi Tof Ms ◽

Maldi Tof ◽

Vineyard Soil ◽

Tof Ms

This study describes a novel fungal species belonging to the genus Gongronella. During a previous work focusing on metalaxyl degradation by Mucorales strains, two isolates from vineyard soil samples collected in the Alentejo region, south Portugal, were identified as a putative novel species based on combined molecular and MALDI-TOF MS data. This new species is described here using a polyphasic approach that combines morphology, internal transcribed spacer of ribosomal DNA (ITS) and 28S ribosomal DNA (LSU) sequence data analysis and proteomic profiling by MALDI-TOF MS. Phenotypic and molecular data enabled this novel species to be clearly distinguished from other Gongronella species with results of combined ITS+LSU analysis showing that the Gongronella species is related to Gongronella butleri and Gongronella brasiliensis. Therefore, from the results of morphological and molecular analyses, isolates MUM 10.262 and MUM 10.263 seem to represent a new Gongronella species and the name Gongronella eborensis sp. nov. is proposed, with the ex-type strain MUM 10.262 (=CCMI 1100=CBS 128763).

Download Full-text

Rapid identification of bacterial and fungal pathogens from positive blood cultures by MALDI-TOF MS

International Journal of Medical Microbiology ◽

10.1016/j.ijmm.2013.03.002 ◽

2013 ◽

Vol 303 (4) ◽

pp. 205-209 ◽

Cited By ~ 43

Author(s):

Christian Leli ◽

Elio Cenci ◽

Angela Cardaccia ◽

Amedeo Moretti ◽

Francesco D’Alò ◽

...

Keyword(s):

Fungal Pathogens ◽

Rapid Identification ◽

Blood Cultures ◽

Maldi Tof Ms ◽

Maldi Tof ◽

Tof Ms

Download Full-text

Identification of Fungi Isolated from Oral Cavity of Patients with HIV Using MALDI–TOF MS

Journal of Clinical Medicine ◽

10.3390/jcm10081570 ◽

2021 ◽

Vol 10 (8) ◽

pp. 1570

Author(s):

Zuzanna Pawlak ◽

Szymon Andrusiów ◽

Magdalena Pajączkowska ◽

Adriana Janczura

Keyword(s):

Fungal Infections ◽

Maldi Tof Ms ◽

Maldi Tof ◽

Flight Mass Spectrometry ◽

Common Mistake ◽

Microbiological Methods ◽

Study Results ◽

Identification Of Fungi ◽

Group A ◽

Tof Ms

Background: A growing incidence of invasive fungal infections, especially among immunocompromised patients, has given increased significance to microbiological diagnostics of yeast-like fungi. More accurate and faster fungi identification methods that can compete with classical methods are being searched for. In this paper, classical microbiological methods are compared to MALDI–TOF MS (matrix-assisted laser desorption/ionization time-of-flight mass spectrometry). Methods: The diagnostic material was collected from buccal mucosa from 98 adults, including 69 with HIV. Only positive cultures were included in the study. Results: Matching results were obtained in 45 samples, and there were nonmatching results in 35 samples, with the majority of these in the study group, constituting 50% of identifications within this group. A particularly common mistake resulting from the use of classical methods is the false identification of C. dubliniensis as C. albicans. Additionally, C. tropicalis proves to be difficult to identify. Conclusions: Our results and literature data suggest that MALDI–TOF MS should be considered an effective alternative to classical methods in terms of fungi identification, especially among HIV-positive patients, due to the different morphology of fungal colonies.

Download Full-text

Use of MALDI TOF-MS Method After Lysis Filtration for Rapid Identification of Yeast

Türk Mikrobiyoloji Cemiyeti Dergisi ◽

10.5222/tmcd.2021.04909 ◽

2021 ◽

Author(s):

Sami Eren ◽

Dilek Yeşim Metin ◽

Süheyla Hilmioğlu Polat

Keyword(s):

Fungal Infections ◽

Rapid Identification ◽

Candida Guilliermondii ◽

Candida Spp ◽

Filtration Method ◽

Maldi Tof Ms ◽

Maldi Tof ◽

School Of Medicine ◽

Reliable Methods ◽

Tof Ms

Objective: The early diagnosis of candidosis is very important in fungal infections and to reduce mortality rates, short-term and reliable methods are needed. In this study, the appropriateness of MALDI-TOF MS method after lysis filtration was investigated for the faster identification of Candida spp. Method: 100 Candida isolated from positive blood cultures sent to Ege University, School of Medicine, Medical Microbiology laboratory, were studied with Dalmau plaque, MALDI-TOF MS and after lysis filtration methods. The results were compared with those obtained from the classical MALDI-TOF MS method. Results: According to conventional and MALDI-TOF MS identification methods; 37 Candida albicans, 23 Candida parapsilosis, 17 Candida tropicalis, nine Candida glabrata, six Candida kefyr, three Candida guilliermondii, three Candida dubliniensis and three Candida krusei were identified. After lysis filtration method; 26 C. albicans, nine C. parapsilosis, 13 C. tropicalis, nine C. glabrata, two C. kefyr, three C. dubliniensis, two C. krusei. MALDI-TOF MS method and Dalmau plaque and after lysis filtration method were found to be compatible in 64 strains. Incompatibility was not due to incorrect identification, but spectrum deficiency. Conclusion: The strains, which are not identified as “spectrum deficiency” consist of the species in the database. Sixty-four Candida species were found to be 100% compatible with the standard methods and this method was found to be advantageous for at least 48 hours compared to MALDI-TOF MS method which required subculture and for at least 72 hours compared to Dalmau plaque method.

Download Full-text

Performance of the new ID-fungi plate using two types of reference libraries (Bruker and MSI) to identify fungi with the Bruker MALDI Biotyper

Medical Mycology ◽

10.1093/mmy/myz138 ◽

2020 ◽

Vol 58 (7) ◽

pp. 946-957 ◽

Cited By ~ 1

Author(s):

Laura Heireman ◽

Sofie Patteet ◽

Sophia Steyaert

Keyword(s):

Growth Medium ◽

Web Application ◽

Fungal Infections ◽

Species Level ◽

Promising Alternative ◽

Maldi Tof Ms ◽

Maldi Tof ◽

Fungal Identification ◽

Significant Difference ◽

Tof Ms

Abstract During the last decade, matrix-assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-TOF MS) has revolutionized the diagnosis of fungal infections. Recently, a new Conidia ID-fungi plate (IDFP) medium was introduced to facilitate growth and sampling of fungi. This study aimed to evaluate the IDFP for fungal MALDI-TOF MS identification by comparison with a standard fungal growth medium using two reference libraries. A total of 75 filamentous fungal isolates (including 32 dermatophytes) were inoculated on IDFP and Sabouraud-gentamicin-chloramphenicol (SGC) agar and identified by MALDI-TOF MS using formic acid/acetonitrile extraction. Both the commercially available Bruker library (version 2.0) and the public available MSI web application (version 2018) were applied. For 15% of the isolates, a faster growth was noticed on IDFP compared to SGC. IDFP enhanced the performance of fungal identification compared to SGC for both MSI (increase of 16% identifications to genus and 5% to species level) and Bruker library (increase of 22% identifications to genus and 8% to species level). In total, only 73% of the tested isolates were present in the Bruker library compared to 92% for MSI library. No significant difference (P = 0.46) in MALDI score between IDFP and SGC was observed for the MSI library, but scores were significantly (P = 0.03) higher for IDFP when using Bruker library, potentially explained by the prevention of agar contamination by using IDFP since the Bruker database was created from liquid media. IDFP is a promising alternative growth medium for MALDI-TOF MS fungal identification which would strongly benefit from optimizing the Bruker reference library.

Download Full-text