A Comprehensive Analysis of MALDI-TOF MS and Ribosomal DNA Sequencing for Identification of Clinical Yeasts

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.

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Abstract A27: Comprehensive analysis of glycans on α1-acid glycoprotein from various cancer patients by MALDI-TOF-MS for assigning a prognostic predictor

10.1158/1940-6207.prev-11-a27 ◽

2011 ◽

Author(s):

Takayuki Asao ◽

Toyo Nishimura ◽

Takashi Hayashi ◽

Hiroyuki Kuwano ◽

Shin Yazawa

Keyword(s):

Cancer Patients ◽

Comprehensive Analysis ◽

Prognostic Predictor ◽

Maldi Tof Ms ◽

Acid Glycoprotein ◽

Maldi Tof ◽

Tof Ms

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Genotyping of Plasmodium falciparum Pyrimethamine Resistance by Matrix-Assisted Laser Desorption-Ionization Time-of-Flight Mass Spectrometry

Antimicrobial Agents and Chemotherapy ◽

10.1128/aac.48.2.466-472.2004 ◽

2004 ◽

Vol 48 (2) ◽

pp. 466-472 ◽

Cited By ~ 9

Author(s):

Florian Marks ◽

Christian G. Meyer ◽

Jürgen Sievertsen ◽

Christian Timmann ◽

Jennifer Evans ◽

...

Keyword(s):

Dna Sequencing ◽

Laser Desorption ◽

Laser Desorption Ionization ◽

Ionization Time ◽

Maldi Tof Ms ◽

Maldi Tof ◽

Flight Mass Spectrometry ◽

Tof Ms ◽

Matrix Assisted Laser ◽

Matrix Assisted Laser Desorption

ABSTRACT Increasing resistance, recrudescences, and treatment failure have led to the replacement of chloroquine with the combination of pyrimethamine (PYR) and sulfadoxine (SDX) as the first-line antimalarial drugs for treatment of uncomplicated Plasmodium falciparum malaria in several areas where this disease is endemic. The development of resistance to PYR-SDX is favored by incomplete treatment courses or by subtherapeutic levels in plasma. PYR-SDX resistance has been associated with several single-nucleotide polymorphisms (SNPs) in the P. falciparum dihydrofolate reductase (pfdhfr) and the P. falciparum dihydropteroate synthetase (pfdhps) genes. We have established assays based on matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) that conveniently allow the identification of SNPs associated with PYR resistance. Variants occurring at codon positions 16, 51, 59, and 108 of the pfdhfr gene were analyzed by MALDI-TOF MS in synthetic oligonucleotides to determine the detection threshold. In addition, 63 blood samples from subjects with P. falciparum parasitemia of various degrees were analyzed. The results were compared to those obtained by DNA sequencing of the respective gene fragment. The results of MALDI-TOF MS and DNA sequencing were consistent in 40 samples. In 23 samples two or three pfdhfr variants were detected by MALDI-TOF assays, whereas DNA-sequencing revealed one variant only. Simultaneous detection of two different mutations by biplex assays was, in principle, feasible. As demonstrated by the example of PYR resistance, MALDI-TOF MS allows for rapid and automated high-throughput assessment of drug sensitivity in P. falciparum malaria.

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Identification of the bacterial population in manganese removal filters

Water Science & Technology Water Supply ◽

10.2166/ws.2016.184 ◽

2016 ◽

Vol 17 (3) ◽

pp. 842-850

Author(s):

Jantinus H. Bruins ◽

Branislav Petrusevski ◽

Yness M. Slokar ◽

Gerhard H. Wübbels ◽

Koen Huysman ◽

...

Keyword(s):

Dna Sequencing ◽

Bacterial Population ◽

Next Generation ◽

Filter Media ◽

Next Generation Dna Sequencing ◽

Maldi Tof Ms ◽

Manganese Removal ◽

Maldi Tof ◽

Ms Analysis ◽

Tof Ms

The aim of this study was to identify bacteria present in ripened manganese removal filters for drinking water production. The bacterial population was identified with ‘next generation’ DNA sequencing, and specific bacteria were quantified with quantitative polymerase chain reaction (qPCR) and characterized by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) analysis. The ‘next generation’ DNA sequencing analysis showed a bacteria population shift from the iron oxidizing species Gallionella spp. in the Fe-filter to manganese and nitrite oxidizing species Pseudomonas spp. and Nitrospira spp., respectively, present in the manganese removal filter. qPCR analysis confirmed the presence of a low concentration of the well-known Mn2+-oxidizing species Ps. putida in the manganese removal filter backwash water. Bacteria of the genus Pseudomonas, isolated from backwash water from a manganese removal filter were cultured and identified with MALDI-TOF MS analysis. Amongst others, P. gessardii, P. grimontii, and P. koreensis were identified. The presence of several manganese oxidizing bacteria species in ripened filter media supports the assumption that a microbial consortium is involved in the oxidation of manganese. Understanding the mechanisms by which manganese coating of filter media commences could endorse the creation of conditions favouring Birnessite formation, and possibly help in reducing typically long ripening periods of manganese removal filters with virgin filter media.

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Identification of polymorphisms within the bovine prion protein gene (Prnp) by DNA sequencing and genotyping by MALDI-TOF-MS

Neurogenetics ◽

10.1007/s10048-001-0126-0 ◽

2002 ◽

Vol 4 (1) ◽

pp. 59-60 ◽

Cited By ~ 20

Author(s):

Andreas Humeny ◽

Katrin Schiebel ◽

Silke Seeber ◽

Cord-Michael Becker

Keyword(s):

Dna Sequencing ◽

Prion Protein ◽

Protein Gene ◽

Prion Protein Gene ◽

Maldi Tof Ms ◽

Maldi Tof ◽

Tof Ms

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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).

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