scholarly journals Identification of bacterial pathogens in cultured fish with a custom peptide database constructed by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS)

2020 ◽  
Author(s):  
Patharapol Piamsomboon ◽  
Janthima Jaresitthikunchai ◽  
Tran Quang Hung ◽  
Sittiruk Roytrakul ◽  
Janenuj Wongtavatchai
Keyword(s):  
Bacterial Pathogens ◽  
Bacterial Species ◽  
Rapid Identification ◽  
Maldi Tof Ms ◽  
Maldi Tof ◽  
Flight Mass Spectrometry ◽  
Custom Made ◽  
Cultured Fish ◽  
Maldi Biotyper ◽  
Tof Ms

Abstract Background: The majority of infectious diseases of cultured fish is caused by bacteria. Rapid identification of bacterial pathogens is necessary for immediate management. The present study developed a custom Main Spectra Profile (MSP) database and validate the method using matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) for rapid identification of fish bacterial pathogens. Streptococcus agalactiae, Streptococcus iniae, Aeromonas hydrophila, Aeromonas veronii, and Edwardsiella tarda obtained from diseased fish were used as representative bacterial pathogens in this study. Bacterial peptides were extracted to create a Main Spectra Profile (MSP), and the MSPs of each bacterial species was added into the MALDI Biotyper database. Fifteen additional isolates of each bacterial species were tested to validate the utilized technique. Results: The MSPs of all field isolates were clearly distinguishable, and the MSPs of the same species were clustered together. The identification methodology was validated with 75 bacterial isolates. The reliability and specificity of the method were determined with MALDI Biotyper log score values and matching results with 16s rDNA sequencing. The species identification using the public MALDI Biotyper library (Bruker MALDI Biotyper) showed unreliable results (log score <2.000) with 42.67% matching result with the reference method. In contrast, accurate identification was obtained when using the custom-made database, giving log score > 2.115, and a 100% matching result. Conclusion: This study demonstrates an effective identification of fish bacterial pathogens when a complete custom-made MSP database is applied. Further applications require a broad, well-established database to accommodate prudent identification of many fish bacterial pathogens by MALDI-TOF MS.

scholarly journals Identification of bacterial pathogens in cultured fish with a custom peptide database constructed by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS)

2020 ◽  
Author(s):  
Patharapol Piamsomboon ◽  
Janthima Jaresitthikunchai ◽  
Tran Quang Hung ◽  
Sittiruk Roytrakul ◽  
Janenuj Wongtavatchai
Keyword(s):  
Bacterial Pathogens ◽  
Bacterial Species ◽  
Rapid Identification ◽  
Ionization Time ◽  
Maldi Tof Ms ◽  
Maldi Tof ◽  
Flight Mass Spectrometry ◽  
Cultured Fish ◽  
Maldi Biotyper ◽  
Tof Ms

Abstract Background: The majority of infectious diseases of cultured fish is caused by bacteria. Rapid identification of bacterial pathogens is necessary for immediate management. The present study developed a matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) for rapid identification of fish bacterial pathogens. Streptococcus agalactiae, Streptococcus iniae, Aeromonas hydrophila, Aeromonas veronii, and Edwardsiella tarda obtained from diseased fish were used as representative bacterial pathogens in this study. Bacterial peptides were extracted to create a Main Spectra Profile (MSP), and the MSPs of each bacterial species was added into the MALDI Biotyper database. Fifteen additional isolates of each bacterial species were tested to validate the utilized technique. Results: The MSPs of all field isolates were clearly distinguishable, and the MSPs of the same species were clustered together. However, the species identification when matched with the public MALDI Biotyper library (Bruker MALDI Biotyper) showed unreliable results. Accurate identification was only obtained when using the custom-made database, giving a 100% matching result with the reference method. Conclusion: This study demonstrates an alternative technique for effective identification of fish bacterial pathogens. Further applications require a broad, well-established database to accommodate prudent identification of many fish bacterial pathogens by MALDI-TOF MS.

scholarly journals Identification of bacterial pathogens in cultured fish using Matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS)

2019 ◽  
Author(s):  
Patharapol Piamsomboon ◽  
Janthima Jaresitthikunchai ◽  
Tran Quang Hung ◽  
Sittiruk Roytrakul ◽  
Janenuj Wongtavatchai
Keyword(s):  
Bacterial Pathogens ◽  
Bacterial Species ◽  
Laser Desorption Ionization ◽  
Ionization Time ◽  
Maldi Tof Ms ◽  
Maldi Tof ◽  
Flight Mass Spectrometry ◽  
Cultured Fish ◽  
Maldi Biotyper ◽  
Tof Ms

Abstract Background Majority of infectious diseases of cultured fish is caused by bacteria. Rapid identification of bacterial pathogen is necessary for immediate management. The present study developed Matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) for a fast identification of fish bacterial pathogens. Streptococcus agalactiae, S. iniae, Aeromonas hydrophila, A. veronii, and Edwardsiella tarda obtained from diseased fish were used as representative bacterial pathogens in this study. Bacterial peptides were extracted to create the Main Spectra Profile (MSP), and the MSP of each bacterial species was added into the MALDI Biotyper database. Ten additional field isolates of each bacterial species were tested to validate the technique.Results The MSP of all field isolates were clearly distinguishable, and the MSPs of the same species were clustered together. However, the species identification when matching with the public MALDI Biotyper library (Bruker MALDI Biotyper) showed unreliable result at the species level. The accurate identification only obtained when the custom-made database was used, giving a 100% matching result.Conclusion This study demonstrates a newly developed technique for the effective identification of fish bacterial pathogens. Further applications require a broad, well-established database to accommodate the prudent identification of many fish bacterial pathogens by MALDI-TOF MS.

scholarly journals Rapid identification of Nocardia cyriacigeorgica from a brain abscess patient using MALDI-TOF-MS

2020 ◽  
Vol 2020 (10) ◽  
Author(s):  
Mohammed AlMogbel ◽  
Mohammed AlBolbol ◽  
Noura Elkhizzi ◽  
Hisham AlAjlan ◽  
John Philip Hays ◽  
...  
Keyword(s):  
Mass Spectrometry ◽  
Bacterial Species ◽  
Rapid Identification ◽  
Clinical Samples ◽  
Maldi Tof Ms ◽  
Maldi Tof ◽  
Flight Mass Spectrometry ◽  
Nocardia Cyriacigeorgica ◽  
Diagnostic Microbiology ◽  
Tof Ms

Abstract Nocardia cyriacigeorgica (N. cyriacigeorgica) is most frequently associated with human infections, including chronic bronchitis, pulmonary disease and brain abscesses. In general, N. cyriacigeorgica causes infections in immunocompromised individuals and has been reported in clinical samples worldwide. However, the isolation and speciation of N. cyriacigeorgica in the routine diagnostic microbiology laboratory are complicated and time consuming. Recent mass spectrometry techniques such as matrix-assisted laser desorption/ionization time-of-flight-mass spectrometry (MALDI-TOF-MS) have been successfully integrated into many routine diagnostic microbiology laboratories, allowing for the rapid, accurate and simple identification and speciation of many different microorganisms, including difficult-to-identify bacterial species. Here, we present a case report of a 65-year-old female patient from the neurology ward of Prince Sultan Military Medical City in Riyadh, Saudi Arabia, who was infected with N. cyriacigeorgica. The bacterium was successfully identified by MALDI-TOF-MS, with species identification subsequently confirmed by sequence analysis of the 16S ribosomal RNA.

scholarly journals Establishment and Application of Matrix-Assisted Laser Desorption/Ionization Time-of-Flight Mass Spectrometry for Detection of Shewanella Genus

2021 ◽  
Vol 12 ◽  
Author(s):  
Keyi Yu ◽  
Zhenzhou Huang ◽  
Ying Li ◽  
Qingbo Fu ◽  
Lirong Lin ◽  
...  
Keyword(s):  
Laser Desorption ◽  
Rapid Identification ◽  
Species Level ◽  
Laser Desorption Ionization ◽  
Ionization Time ◽  
Maldi Tof Ms ◽  
Maldi Tof ◽  
Flight Mass Spectrometry ◽  
Type Strains ◽  
Tof Ms

Shewanella species are widely distributed in the aquatic environment and aquatic organisms. They are opportunistic human pathogens with increasing clinical infections reported in recent years. However, there is a lack of a rapid and accurate method to identify Shewanella species. We evaluated here matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) for rapid identification of Shewanella. A peptide mass reference spectra (PMRS) database was constructed for the type strains of 36 Shewanella species. The main spectrum projection (MSP) cluster dendrogram showed that the type strains of Shewanella species can be effectively distinguished according to the different MS fingerprinting. The PMRS database was validated using 125 Shewanella test strains isolated from various sources and periods; 92.8% (n = 116) of the strains were correctly identified at the species level, compared with the results of multilocus sequence analysis (MLSA), which was previously shown to be a method for identifying Shewanella at the species level. The misidentified strains (n = 9) by MALDI-TOF MS involved five species of two groups, i.e., Shewanella algae–Shewanella chilikensis–Shewanella indica and Shewanella seohaensis–Shewanella xiamenensis. We then identified and defined species-specific biomarker peaks of the 36 species using the type strains and validated these selected biomarkers using 125 test strains. Our study demonstrated that MALDI-TOF MS was a reliable and powerful tool for the rapid identification of Shewanella strains at the species level.

scholarly journals Evaluation of matrix-assisted laser desorption ionization-time of flight mass spectrometry in diagnosis of clinical Nocardia species

2019 ◽  
Vol 43 (3) ◽  
pp. 157-162
Author(s):  
Gülşen Hasçelik ◽  
Markus Kostrzewa ◽  
Hamit Kaan Müştak ◽  
Celalettin Uner ◽  
Kadir Serdar Diker
Keyword(s):  
Species Level ◽  
Rrna Gene ◽  
Laser Desorption Ionization ◽  
Ionization Time ◽  
Maldi Tof Ms ◽  
Maldi Tof ◽  
Flight Mass Spectrometry ◽  
Maldi Biotyper ◽  
Rrna Gene Sequencing ◽  
Tof Ms

Abstract Background The routine identification to the species level of Nocardia genus by conventional methods is a fastidious and time-consuming process owing to the limited biochemical reactivity of these microorganisms, often requiring 1 or more days to complete identification. Matrix-assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF MS) is a new technology for definitive and rapid species identification. Methods We evaluated the MALDI-TOF MS for the identification of 44 clinical isolates of Nocardia species in comparison to 16S ribosomal RNA (rRNA) gene sequencing. Nocardia isolates were identified by microbiological examination, phenotypical tests and MALDI-TOF MS and the results were compared by 16S rRNA gene sequencing. Results Of the 44 Nocardia strains, the identification of 28 isolates was determined with MALDI Biotyper database. According to this, 16 isolates (57.1%) of the strain log scores were ≥2. Two (7.1%) were identified to the species level (log scores of ≥2) as Nocardia otitidiscaviarum. The addition of a newly established Nocardia database (16 new Nocardia strains included to the original database) did significantly improve the scores. The results were 43 (97.7%) correct identification to the species level (log scores of ≥2). Conclusions This study showed that the identification of clinical Nocardia isolates by the Bruker MALDI Biotyper is highly reliable, whereas identification rates are generally lower than those for some Gram-negative bacteria and Gram-positive cocci. Based on our data, the identification rates can be improved by validated new database entries and the results can be confirmed with nucleic acid sequence analysis.

scholarly journals Evaluation of Matrix-Assisted Laser Desorption/Ionization Time-of-Flight Mass Spectrometry for Rapid Identification of Bacteria in Processed Soybean Products

2013 ◽  
Vol 2 (3) ◽  
pp. 104 ◽  
Author(s):  
Yuko Furukawa ◽  
Mitsuru Katase ◽  
Kazunobu Tsumura
Keyword(s):  
Laser Desorption ◽  
Rapid Identification ◽  
Bacterial Cells ◽  
Laser Desorption Ionization ◽  
Ionization Time ◽  
Maldi Tof Ms ◽  
Maldi Tof ◽  
Flight Mass Spectrometry ◽  
Identification Of Bacteria ◽  
Tof Ms

Matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) has recently been demonstrated as a rapid and reliable method for identifying bacteria in colonies grown on culture plates. Rapid identification of food spoilage bacteria is important for ensuring the quality and safety of food. To shorten the time of analysis, several researchers have proposed the direct MALDI-TOF MS tequnics for identification of bacteria in clinical samples such as urine and positive blood cultures. In this study, processed soybean products (total 26 test samples) were initially conducted a culture enrichiment step and bacterial cells were separated from interfering components. Harvested bacterial cells were determined by MALDI-TOF MS and 16S rRNA gene sequencing method. Six processed soybean products (23%) were increased bacterial cells after culture enrichiment step and they were sucessfully obtained the accurate identification results by MALDI-TOF MS-based method without colony formation.

scholarly journals Rapid identification of Legionella species by mass spectrometry

2010 ◽  
Vol 59 (3) ◽  
pp. 273-284 ◽  
Author(s):  
Claire Moliner ◽  
Christophe Ginevra ◽  
Sophie Jarraud ◽  
Christophe Flaudrops ◽  
Marielle Bedotto ◽  
...  
Keyword(s):  
Mass Spectrometry ◽  
Bacterial Species ◽  
Rapid Identification ◽  
Species Level ◽  
Blind Test ◽  
Maldi Tof Ms ◽  
Maldi Tof ◽  
Tract Infections ◽  
Pcr Targeting ◽  
Tof Ms

Legionella species are facultative, intracellular bacteria that infect macrophages and protozoa, with the latter acting as transmission vectors to humans. These fastidious bacteria mostly cause pulmonary tract infections and are routinely identified by various molecular methods, mainly PCR targeting the mip gene and sequencing, which are expensive and time-consuming. Recently, matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF-MS) has emerged as a rapid and inexpensive method for identification of bacterial species. This study evaluated the use of MALDI-TOF-MS for rapid species and serogroup identification of 21 Legionella species recognized as human pathogens. To this end, a reference MS database was developed including 59 Legionella type strains, and a blind test was performed using 237 strains from various species. Two hundred and twenty-three of the 237 strains (94.1 %) were correctly identified at the species level, although ten (4.2 %) were identified with a score lower than 2.0. Fourteen strains (5.9 %) from eight species were misidentified at the species level, including seven (3.0 %) with a significant score, suggesting an intraspecific variability of protein profiles within some species. MALDI-TOF-MS was reproducible but could not identify Legionella strains at the serogroup level. When compared with mip gene sequencing, MALDI-TOF-MS exhibited a sensitivity of 99.2 and 89.9 % for the identification of Legionella strains at the genus and species level, respectively. This study demonstrated that MALDI-TOF-MS is a reliable tool for the rapid identification of Legionella strains at the species level.

scholarly journals MDRSA: A Web Based-Tool for Rapid Identification of Multidrug Resistant Staphylococcus aureus Based on Matrix-Assisted Laser Desorption Ionization-Time of Flight Mass Spectrometry

2021 ◽  
Vol 12 ◽  
Author(s):  
Chia-Ru Chung ◽  
Zhuo Wang ◽  
Jing-Mei Weng ◽  
Hsin-Yao Wang ◽  
Li-Ching Wu ◽  
...  
Keyword(s):  
Staphylococcus Aureus ◽  
Rapid Identification ◽  
Antibiotics Resistance ◽  
Web Tool ◽  
Web Based ◽  
Ionization Time ◽  
Maldi Tof Ms ◽  
Maldi Tof ◽  
Flight Mass Spectrometry ◽  
Tof Ms

As antibiotics resistance on superbugs has risen, more and more studies have focused on developing rapid antibiotics susceptibility tests (AST). Meanwhile, identification of multiple antibiotics resistance on Staphylococcus aureus provides instant information which can assist clinicians in administrating the appropriate prescriptions. In recent years, matrix-assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF MS) has emerged as a powerful tool in clinical microbiology laboratories for the rapid identification of bacterial species. Yet, lack of study devoted on providing efficient methods to deal with the MS shifting problem, not to mention to providing tools incorporating the MALDI-TOF MS for the clinical use which deliver the instant administration of antibiotics to the clinicians. In this study, we developed a web tool, MDRSA, for the rapid identification of oxacillin-, clindamycin-, and erythromycin-resistant Staphylococcus aureus. Specifically, the kernel density estimation (KDE) was adopted to deal with the peak shifting problem, which is critical to analyze mass spectra data, and machine learning methods, including decision trees, random forests, and support vector machines, which were used to construct the classifiers to identify the antibiotic resistance. The areas under the receiver operating the characteristic curve attained 0.8 on the internal (10-fold cross validation) and external (independent testing) validation. The promising results can provide more confidence to apply these prediction models in the real world. Briefly, this study provides a web-based tool to provide rapid predictions for the resistance of antibiotics on Staphylococcus aureus based on the MALDI-TOF MS data. The web tool is available at: http://fdblab.csie.ncu.edu.tw/mdrsa/.

scholarly journals Rapid and cost-effective identification of Bartonella species using mass spectrometry

2009 ◽  
Vol 58 (9) ◽  
pp. 1154-1159 ◽  
Author(s):  
Pierre-Edouard Fournier ◽  
Carine Couderc ◽  
Sylvain Buffet ◽  
Christophe Flaudrops ◽  
Didier Raoult
Keyword(s):  
Mass Spectrometry ◽  
Species Identification ◽  
Chemical Reactivity ◽  
Bacterial Species ◽  
Cost Effective ◽  
Molecular Techniques ◽  
Maldi Tof Ms ◽  
Maldi Tof ◽  
Flight Mass Spectrometry ◽  
Tof Ms

Bacteria of the genus Bartonella are emerging zoonotic bacteria recognized in a variety of human diseases. Due to their poor chemical reactivity, these fastidious bacteria are poorly characterized using routine phenotypic laboratory tests. Identification is usually achieved using molecular techniques that are time-consuming, expensive and technically demanding. Recently, matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) has emerged as a new technique for bacterial species identification. This study evaluated the use of MALDI-TOF MS for rapid genus and species identification of Bartonella species. Reference strains representing 17 recognized Bartonella species were studied. For each species, MS spectra for four colonies were analysed. The consensus spectrum obtained for each species was unique among spectra obtained for 2843 bacteria within the Bruker database, including 109 alphaproteobacteria. Thirty-nine additional blind-coded Bartonella strains were correctly identified at the species level, including 36 with a significant score. Altogether, these data demonstrate that MS is an accurate and reproducible tool for rapid and inexpensive identification of Bartonella species.

scholarly journals Rapid Identification of Vibrio parahaemolyticus by Whole-Cell Matrix-Assisted Laser Desorption Ionization-Time of Flight Mass Spectrometry

2009 ◽  
Vol 75 (21) ◽  
pp. 6745-6756 ◽  
Author(s):  
Tracy H. Hazen ◽  
Robert J. Martinez ◽  
Yanfeng Chen ◽  
Patricia C. Lafon ◽  
Nancy M. Garrett ◽  
...  
Keyword(s):  
Vibrio Parahaemolyticus ◽  
Laser Desorption ◽  
Rapid Identification ◽  
Laser Desorption Ionization ◽  
Ionization Time ◽  
Whole Cell ◽  
Maldi Tof Ms ◽  
Maldi Tof ◽  
Flight Mass Spectrometry ◽  
Tof Ms

ABSTRACT Vibrio parahaemolyticus is a pathogenic marine bacterium that is the main causative agent of bacterial seafood-borne gastroenteritis in the United States. An increase in the frequency of V. parahaemolyticus-related infections during the last decade has been attributed to the emergence of an O3:K6 pandemic clone in 1995. The diversity of the O3:K6 pandemic clone and its serovariants has been examined using multiple molecular techniques including multilocus sequence analysis, pulsed-field gel electrophoresis, and group-specific PCR analysis. Matrix-assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF MS) has become a powerful tool for rapidly distinguishing between related bacterial species. In the current study, we demonstrate the development of a whole-cell MALDI-TOF MS method for the distinction of V. parahaemolyticus from other Vibrio spp. We identified 30 peaks that were present only in the spectra of the V. parahaemolyticus strains examined in this study that may be developed as MALDI-TOF MS biomarkers for identification of V. parahaemolyticus. We detected variation in the MALDI-TOF spectra of V. parahaemolyticus strains isolated from different geographical locations and at different times. The MALDI-TOF MS spectra of the V. parahaemolyticus strains examined were distinct from those of the other Vibrio species examined including the closely related V. alginolyticus, V. harveyi, and V. campbellii. The results of this study demonstrate the first use of whole-cell MALDI-TOF MS analysis for the rapid identification of V. parahaemolyticus.

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