Dataset Information

Proteotyping of Campylobacter jejuni by MALDI-TOF MS and Strain Solution Version 2 Software.

ABSTRACT: Identification of microorganisms by MALDI-TOF MS has become a popular method in the past 20 years. Strain Solution ver. 2 software appended with MALDI-TOF MS enables accurate discrimination of serotypes and strains beyond the genus and species level by creating a theoretical mass-based database. In this study, we constructed a theoretical mass database with the validated biomarkers to proteotype Campylobacter jejuni. Using 10 strains belonging to Campylobacter spp. available from culture collections and 41 Campylobacter jejuni strains isolated from humans and foods, the ribosomal protein subunits L36, L32, S14, L24, L23, L7/L12, and S11 could be selected as the effective biomarkers for the proteotyping of C. jejuni at MALDI-TOF MS. An accurate database of their theoretical mass-based values was constructed by matching these gene DNA sequences and the observed mass peaks. We attempted to automatically classify 41 strains isolated from nature using this database and Strain Solution ver. 2 software, and 38 strains (93%) were correctly classified into the intended group based on the theoretical mass-based values. Thus, the seven biomarkers found in this study and Strain Solution ver. 2 are promising for the proteotyping of C. jejuni by MALDI-TOF MS.

SUBMITTER: Ojima-Kato T

PROVIDER: S-EPMC9866418 | biostudies-literature | 2023 Jan

REPOSITORIES: biostudies-literature

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Proteotyping of Campylobacter jejuni by MALDI-TOF MS and Strain Solution Version 2 Software.

Ojima-Kato Teruyo T Nagai Satomi S Fujita Akane A Sakata Junko J Tamura Hiroto H

Microorganisms 20230112 1

Identification of microorganisms by MALDI-TOF MS has become a popular method in the past 20 years. Strain Solution ver. 2 software appended with MALDI-TOF MS enables accurate discrimination of serotypes and strains beyond the genus and species level by creating a theoretical mass-based database. In this study, we constructed a theoretical mass database with the validated biomarkers to proteotype Campylobacter jejuni. Using 10 strains belonging to Campylobacter spp. available from c ...[more]

PMID: 36677494

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Project description:Elizabethkingia species, isolated from clinical and environmental samples, are emerging opportunistic bacterial pathogens with a high mortality rate in clinical settings worldwide. Taxonomically, Elizabethkingia comprises seven species: E. anophelis, E. argenteiflava, E. bruuniana, E. meningoseptica, E. miricola, E. ursingii, and E. occulta. In this study, we identified useful biomarker proteins, including ribosomal L29, L30, S21, and the YtxH domain-containing proteins, for distinguishing Elizabethkingia species using matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) profiles. Evaluation of 29 clinical and environmental Elizabethkingia strains revealed that these species could be separated by MALDI-TOF MS profiles into six groups-E. anophelis, E. argenteiflava, E. bruuniana/E. miricola, E. meningoseptica, E. ursingii, and E. occulta-based on the four biomarker protein peaks. This study demonstrates the potential of routine MALDI-TOF MS -based examination methods for the early detection of Elizabethkingia species in clinical laboratories.ImportanceElizabethkingia species are groups of emerging opportunistic bacterial pathogens with a high mortality rate, causing healthcare-associated outbreaks worldwide. Rapid identification of Elizabethkingia species is important becausethese species show intrinsically carbapenem resistance and there are few data for using appropriate antibiotics. Until now, only whole-genome sequencing could accurately identify the seven Elizabethkingia species. Therefore, establishing rapid and accurate identification methods for Elizabethkingia species in clinical laboratories is vital. In this study, we developed new methods for identifying Elizabethkingia species using four biomarker protein peaks-ribosomal L29, L30, S21, and the YtxH domain-containing proteins by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) proteotyping. This study demonstrates the potential of routine MALDI-TOF MS -based laboratory examination for the early identification of Elizabethkingia species.

Project description:BackgroundIn the last decade, an innovative approach has emerged for arthropod identification based on matrix-assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-TOF MS). Increasing interest in applying the original technique for arthropod identification has led to the development of a variety of procedures for sample preparation and selection of body parts, among others. However, the absence of a consensual strategy hampers direct inter-study comparisons. Moreover, these different procedures are confusing to new users. Establishing optimized procedures and standardized protocols for mosquito identification by MALDI-TOF MS is therefore a necessity, and would notably enable the sharing of reference MS databases. Here, we assess the optimal conditions for mosquito identification using MALDI-TOF MS profiling.MethodsThree homogenization methods, two of which were manual and one automatic, were used on three distinct body parts (legs, thorax, head) of two mosquito laboratory strains, Anopheles coluzzii and Aedes aegypti, and the results evaluated. The reproducibility of MS profiles, identification rate with relevant scores and the suitability of procedures for high-throughput analyses were the main criteria for establishing optimized guidelines. Additionally, the consequences of blood-feeding and geographical origin were evaluated using both laboratory strains and field-collected mosquitoes.ResultsRelevant score values for mosquito identification were obtained for all the three body parts assayed using MALDI-TOF MS profiling; however, the thorax and legs were the most suitable specimens, independently of homogenization method or species. Although the manual homogenization methods were associated with a high rate of identification on the three body parts, this homogenization mode is not adaptable to the processing of a large number of samples. Therefore, the automatic homogenization procedure was selected as the reference homogenization method. Blood-feeding status did not hamper the identification of mosquito species, despite the presence of MS peaks from original blood in the MS profiles of the three body parts tested from both species. Finally, a significant improvement in identification scores was obtained for field-collected specimens when MS spectra of species from the same geographical area were added to the database.ConclusionThe results of the current study establish guidelines for the selection of mosquito anatomic parts and modality of sample preparation (e.g. homogenization) for future specimen identification by MALDI-TOF MS profiling. These standardized operational protocols could be used as references for creating an international MS database.

Dataset Information

Proteotyping of Campylobacter jejuni by MALDI-TOF MS and Strain Solution Version 2 Software.

Publications

Proteotyping of <i>Campylobacter jejuni</i> by MALDI-TOF MS and Strain Solution Version 2 Software.

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