• Mass Spectrometry Letters
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• v.5 no.2
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• pp.49-51
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• 2014

The effectiveness of tertiary matrices composed of the combination of three common matrices (dihydrobenzoic acid (DHB), ${\alpha}$-cyano-4-hydroxycinnamic acid (CHCA), and sinapinic acid (SA)) was compared with that of single or binary matrices in the analysis of polyethylene glycol (PEG) polymers ranging from 1400 to 10000 Da using matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS). A tertiary matrix of 2,5-DHB+CHCA+SA was the most effective in terms of S/N ratios. CHCA and CHCA+SA produced the highest S/N ratios among the single matrices and the binary matrices, respectively. The improvement observed when using a tertiary matrix in analyses of PEG polymers by MALDI-TOF MS is believed to be due to the uniform morphology of the MALDI sample spots and synergistic effects arising from the mixture of the three matrix materials.

• Bulletin of the Korean Chemical Society
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• v.20 no.11
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• pp.1299-1302
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• 1999

Matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF-MS) was used to demonstrate cross-linking of peptides induced by transglutaminase. The presence of ε-( Υ-glutamyl)lysine isopeptide cross-link in the acid hydrolysate of the cross-linking reaction mixture was also demonstrated by MALDI-TOF-MS without prior separation. MALDI-TOF-MS quickly provided peptide mass maps after pronase digestion of the cross-linked peptide adduct, which enabled us to monitor the hydrolytic sequence. Pronase appears to preferentially hydrolyze peptide bonds distant from the cross-link before hydrolyzing peptide bonds around the cross-link. The results suggest that pronase digestion followed by MALDI-TOF-MS could be used for determination of amino acid sequence around a modification site.

• Korean Journal of Clinical Laboratory Science
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• v.55 no.4
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• pp.244-252
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• 2023

The timeliness and accuracy of test results are crucial factors for clinicians to decide and promptly administer effective and targeted antimicrobial therapy, especially in life-threatening infections or when vital organs and functions, such as sight, are at risk. Further research is needed to refine and optimize matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS)-based assays to obtain accurate and reliable results in the shortest time possible. MALDI-TOF MS-based bacterial identification focuses primarily on techniques for isolating and purifying pathogens from clinical samples, the expansion of spectral libraries, and the upgrading of software. As technology advances, many MALDI-based microbial identification databases and systems have been licensed and put into clinical use. Nevertheless, it is still necessary to develop MALDI-TOF MS-based antimicrobial-resistance analysis for comprehensive clinical microbiology characterization. The important applications of MALDI-TOF MS in clinical research include specific application categories, common analytes, main methods, limitations, and solutions. In order to utilize clinical microbiology laboratories, it is essential to secure expertise through education and training of clinical laboratory scientists, and database construction and experience must be maximized. In the future, MALDI-TOF mass spectrometry is expected to be applied in various fields through the use of more powerful databases.

• Journal of Microbiology and Biotechnology
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• v.29 no.4
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• pp.548-557
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• 2019

Staphylococcus species have a ubiquitous habitat in a wide range of foods, thus the ability to identify staphylococci at the species level is critical in the food industry. In this study, we performed rapid identification of Staphylococcus species using Matrix-Assisted Laser Desorption/Ionization Time-of-Flight mass spectrometry (MALDI-TOF MS). MALDI-TOF MS was evaluated for the identification of Staphylococcus reference strains (n = 19) and isolates (n = 96) from various foods with consideration for the impact of sample preparation methods and incubation period. Additionally, the spectra of isolated Staphylococcus strains were analyzed using principal component analysis (PCA) and a main spectra profile (MSP)-based dendrogram. MALDI-TOF MS accurately identified Staphylococcus reference strains and isolated strains: the highest performance was by the EX method (83.3~89.5% accuracy) at species level identification (EDT, 70.3~78.9% accuracy; DT, less than 46.3~63.2% accuracy) of 24-h cultured colonies. Identification results at the genus level were 100% accurate at EDT, EX sample preparation and 24-h incubation time. On the other hand, the DT method showed relatively low identification accuracy in all extraction methods and incubation times. The analyzed spectra and MSP-based dendrogram showed that the isolated Staphylococcus strains were characterized at the species level. The performance analysis of MALDI-TOF MS shows the method has the potential ability to discriminate between Staphylococcus species from foods in Korea. This study provides valuable information that MALDI-TOF MS can be applied to monitor microbial populations and pathogenic bacteria in the food industry thereby contributing to food safety.

• Korean Journal of Clinical Laboratory Science
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• v.50 no.4
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• pp.414-421
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• 2018

In this study, Gram-positive bacilli (GPB) were identified by MALDI-TOF MS and analyzed according to the quaternary and microbial strains in the blood culture medium over a two year period at a university hospital. The results were as follows. The overall positive rate of blood culture was 9.97%. In 713 isolated GPB, 410 strains (57.5%) were identified using a microflex MALDI Biotyper. The positive rate of GPB among the blood culture positive bacteria was 8.2%, and the quarterly isolation rate was 9.8% in the third quarter of 2015, 8.7% in the second quarter of 2016, 8.1% in the third quarter of 2016, 8.1% in the first quarter of 2015, 7.9% in the first quarter of 2015, 7.9% in the second quarter of 2015, 6.8% in the first quarter of 2016, and 6.7% in the fourth quarter of 2015. The isolates were Corynebacterium striatum 89 (12.4%), Bacillus cereus 60 (8.4%), Bacillus subtilis 30 (4.2%), Paenibacillus urinalis 29 (4.1%), and Listeria monocytogenes 25 (3.5%). The results of 16S rRNA sequencing of 43 isolates (86.0%) were consistent with those of the other 50 isolates. Five out of the seven unmatched weeks were not identified by MALDI-TOF MS.

• Journal of Microbiology and Biotechnology
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• v.27 no.6
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• pp.1128-1137
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• 2017

The aim of this study was to explore the accuracy and feasibility of matrix-assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-TOF MS) in identifying bacteria from environmental sources, as compared with rpoA gene sequencing, and to evaluate the occurrence of bacteria of the genus Enterococcus in wild birds. In addition, a phyloproteomic analysis of certain Enterococcus species with spectral relationships was performed. The enterococci were isolated from 25 species of wild birds in central Europe (Poland). Proteomic (MALDI-TOF MS) and genomic (rpoA gene sequencing) methods were used to identify all the isolates. Using MALDI-TOF MS, all 54 (100%) isolates were identified as Enterococcus spp. Among these, 51 (94.4%) isolates were identified to the species level (log(score) ${\geq}2.0$), and three isolates (5.6%) were identified at a level of probable genus identification (log(score) 1.88-1.927). Phylogenetic analysis based on rpoA sequences confirmed that all enterococci had been correctly identified. Enterococcus faecalis was the most prevalent enterococcal species (50%) and Enterococcus faecium (33.3%) the second most frequent species, followed by Enterococcus hirae (9.3%), Enterococcus durans (3.7%), and Enterococcus casseliflavus (3.7%). The phyloproteomic analysis of the spectral profiles of the isolates showed that MALDI-TOF MS is able to differentiate among similar species of the genus Enterococcus.

• Korean Journal of Food Science and Technology
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• v.48 no.5
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• pp.437-444
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• 2016

Rapid and reliable identification of microorganisms is important to maintain food quality and to control safety. MALDI-TOF MS-based identification methods are relatively fast and simple compared to other conventional methods including gram staining and biochemical characterization. A colony on subcultured media can be directly prepared on the analysis plate without further complex treatments. In this study, we confirmed the applicability of MALDI-TOF MS-based identification of foodborne pathogens such as Salmonella Enteritidis/Typhimurium, Staphylococcus aureus, Vibrio parahaemolyticus, Clostridium perfringens, Listeria monocytogenes, Yersinia enterocolitica, Bacillus cereus, Campylobacter jejuni, Campylobacter coli, and Cronobacter sakazakii on the Korea Food Standard Codex. MALDI-TOF MS data of the pathogenic reference strains were incorporated into a commercial MicroID (ASTA Inc.) database. Other pathogenic reference strains and seven isolates from various food samples were correctly identified to the species level by using the MicroID database. In conclusion, MALDI-TOF MS is comparable with commercial biochemical identification.

• Journal of Microbiology and Biotechnology
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• v.25 no.9
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• pp.1537-1541
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• 2015

Previous detection methods for Citrobacter are considered time consuming and laborious. In this study, we have developed a rapid and accurate detection method for Citrobacter species in pork products, using matrix-assisted laser desorption ionization time-of-flight (MALDI-TOF) mass spectrometry (MS). A total of 35 Citrobacter strains were isolated from 30 pork products and identified by both MALDI-TOF MS and 16S rRNA gene sequencing approaches. All isolates were identified to the species level by the MALDI-TOF MS, while 16S rRNA gene sequencing results could not discriminate them clearly. These results confirmed that MALDITOF MS is a more accurate and rapid detection method for the identification of Citrobacter species.

• Mass Spectrometry Letters
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• v.9 no.2
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• pp.56-60
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• 2018

In this study, matrix-assisted laser desorption/ionization (MALDI) was applied to the TEMPO-assisted FRIPS for the first time. We found that 3-HPA is the optimal matrix for the analysis of p-TEMPO-Bz-Sc-peptides, which gives minimal precursor fragmentations. MALDI-TOF/TOF experiments on p-TEMPO-Bz-Sc-peptides yielded mainly $[a_n+H]^+$, $[z_n+H]^+$, and $[y_n]^+-type$ products, indicating that radical-driven peptide fragmentation occurs in MALDI-TOF/TOF-MS.

• Bulletin of the Korean Chemical Society
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• v.32 no.8
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• pp.2827-2829
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• 2011