• 대한임상검사과학회지
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• 제50권4호
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• pp.414-421
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• 2018

본 연구에서는 일개 대학병원에서 2년간 분리된 혈액배양 양성배지에서 MALDI-TOF MS system을 이용하여 Gram-positive bacilli를 동정한 결과를 균종별, 분기별로 분석하였다. Corynebacterium striatum은 총 89균주 중 66균주 (74.2%), Bacillus cereus는 60균주 중 44 균주 (73.3%), Listeria monocytogenes는 25균주 중 25균주 (100%)로 2.0이상의 높은 스코어에서 동정되었다. 미 동정 된 균주는 303균주 중 293균주는 혈액배양에서 1회 분리 균주로 감염균으로서의 의의가 없는 오염 균주로 간주되었다. 감염균으로서 의의가 있는 동일 환자 2회 이상 분리 균주 대상 16S-rRNA sequencing 비교결과 총 50균주 중 43균주가 일치해 86.0% 동정이 가능하였다. 일치하지 않은 7균주 중 5균주는 MALDI-TOF MS로도 동정이 되지 않았다. 결론적으로 혈액배양에서 Gram-positive bacilli가 동정되는 경우, 일차적으로 MALDI-TOF MS를 이용하여 동정해보고 이를 활용한다면 어렵고 비용이 많이 들던 Gram-positive bacilli 동정이 저비용으로 더욱 간편하고 정확해지며, Gram-positive bacilli에 의한 감염 진단에도 도움이 될 것으로 판단된다.

• Journal of Microbiology and Biotechnology
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• 제27권6호
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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.

• Journal of Microbiology and Biotechnology
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• 제29권4호
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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.

• Bulletin of the Korean Chemical Society
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• 제32권2호
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• pp.477-482
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• 2011

The structural analysis of polytrimethylene terephthalate (PTT) and characterization of the hydrolytic degradation products after acid hydrolysis were performed using MALDI-TOF mass spectrometry. Mass spectra of the PTT samples were analyzed using a self-calibration method as well as an internal calibration method with standard materials of known masses. PTT structures constituting the samples were determined from the analyses of the spectra, and their relative compositions were estimated. The MALDI-TOF mass spectra of the acid-hydrolyzed PTT sample showed three main series of oligomer products with different end groups in accordance with the hydrolysis schemes. From the spectra of both $Na^+$ and $K^+$ adducts, it was concluded that the PTT samples have higher affinity for $Na^+$ compared with $K^+$ and therefore show higher ionization efficiency with sodium ions when dithranol is used as a matrix. Two different wavelength laser beams ($\lambda$ = 337 nm and 355 nm) were tested and it was observed that the 355 nm beam was more efficient in obtaining the MALDI spectra of PTT using dithranol as a matrix under our experimental conditions.

• Bulletin of the Korean Chemical Society
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• 제32권8호
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• pp.2827-2829
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• 2011

• Bulletin of the Korean Chemical Society
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• 제33권3호
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• pp.833-838
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• 2012

The thermal degradation products of polytrimethylene terephthalate (PTT) obtained by heating the sample in the temperature range of $250-360^{\circ}C$ under non-oxidative conditions was characterized using MALDI-TOF (matrix assisted laser desorption/ionization) mass spectrometry. The structures of the degradation products were determined and the relative compositions were estimated. The MALDI-TOF mass spectra of the thermally degraded PTT sample showed three main series of oligomer products with different end groups, which were carboxyl/carboxyl, carboxyl/allyl, and allyl/allyl. In contrast to the thermal degradation of polyethylene terephthalate (PET), the oligomers containing terephthalic anhydrides were not detected, whereas the formation of oligomers containing the unsaturated allyl ester group was confirmed by mass assignment. From these results, it was concluded that the thermal degradation of PTT proceeds exclusively through the ${\beta}$-CH hydrogen transfer mechanism, which is in accordance with the proposed reaction mechanism for the thermal degradation of polybutylene terephthalate (PBT).

• Journal of Microbiology and Biotechnology
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• 제25권9호
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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.

• 대한전기학회:학술대회논문집
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• 대한전기학회 2008년도 제39회 하계학술대회
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• pp.1466-1467
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• 2008

We describe here a novel micro total analysis system for the purification and identification of the affinity-captured proteins. Also we demonstrated the mass analysis of the Carcinoembrionic antigen (CEA) and Alpha femtoprotein which were chosen as the target cancer marker. For MALDI-TOF analyses, the proteins should to be separated from a protein mixture and be concentrated when needed. This procedure usually takes a long time even before protease-digested samples are to be obtained from them. Here, we describe integrated and efficient micro chip for protein purification and digestion for MALDI-TOF analyses. At first, disease protein is purified by passing the micro chamber from a protein mixture or human whole serum and released from the micro affinity beads by thermal heating. Purified protein is then transfer to the hole for trypsin digestion. The final sample is analyzed by MALDI-TOF. All the processes could be finished successfully within one hour, which renders MALDI-TOF analyses of a target protein quite simple.

• 한국동물위생학회지
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• 제41권2호
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• pp.105-110
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• 2018

A total of 41 Salmonella (S.) strains were isolated from pigs suffered with severe watery diarrhea and were tried to identify by both matrix-assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF MS) and polymerase chain reaction (PCR) analysis. Fibrinous exudate and ulceration in the large intestine were prevalent in gross observation, and variable degrees of enteritis were observed in the histology of large intestines. Subsequent polymerase chain reaction (PCR) analyses demonstrated that 41 strains were identified as S. Typhimurium (39 strains), though 2 stains were failed to identify. Further identification was performed using both direct smear and protein extraction method by MALDI-TOF MS analyses. In terms of extraction methods, 100% (41/41) of isolates were identified to species level of S. spp. Whereas only 43.9% (18/41) were identified to species level using the direct method. These results thus suggest that rapid and accurate diagnosis of porcine salmonellosis can be guaranteed by MALDI-TOF MS combined with protein extraction method.

• Bulletin of the Korean Chemical Society
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• 제25권12호
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• pp.1791-1800
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• 2004

Angiotensin I, a model decapeptide, was glycosylated and partially hydrolyzed with HCl (6 N, 80 $^{\circ}C$, 4 h), aminopeptidase, and carboxypeptidase Y. A single peptide mass map obtained from truncated peptides in the partial acid hydrolysate of angiotensin and its glycosylation product mixture by matrix-assisted laser desorption/ionization time-of-flight (MALDI-TOF) mass spectrometry enabled sequencing of angiotensin by a combinatorial procedure. MALDI-TOF and electrospray ionization (ESI) tandem mass spectrometric results indicate that both the N-terminal amino group of aspartic acid and the guanidinium group of the second residue arginine are glycosylated.