DNA-Mediated rolling circle amplification for ultrasensitive detection of thrombin using MALDI-TOF mass spectrometry

2018 ◽  
Vol 54 (82) ◽  
pp. 11546-11549 ◽  
Author(s):  
Zengnan Wu ◽  
Ling Lin ◽  
Mashooq Khan ◽  
Weifei Zhang ◽  
Sifeng Mao ◽  
...  
Keyword(s):  
Mass Spectrometry ◽  
Rolling Circle Amplification ◽  
Ultrasensitive Detection ◽  
Specific Detection ◽  
Rolling Circle ◽  
Maldi Tof Ms ◽  
Maldi Tof Mass Spectrometry ◽  
Maldi Tof ◽  
Tof Ms

A DNA-mediated rolling circle amplification (RCA) strategy was established for ultrasensitive and specific detection of thrombin via MALDI-TOF MS.

scholarly journals The clinical relevance of pathogen Aerococcus urinae identification by MALDI-TOF mass spectrometry: a case of sepsis in 86-year-old Caucasian male

2020 ◽  
Vol 35 (2) ◽  
Author(s):  
Jari Intra ◽  
Cecilia Sarto ◽  
Giuseppe Serra ◽  
Paolo Brambilla
Keyword(s):  
Mass Spectrometry ◽  
Urinary Tract ◽  
Previous History ◽  
Blood Stream ◽  
Maldi Tof Ms ◽  
Maldi Tof Mass Spectrometry ◽  
Maldi Tof ◽  
Microbiological Methods ◽  
Aerococcus Urinae ◽  
Tof Ms

The infrequency of urinary tract and blood stream infections caused by Aerococcus urinae is most probably due to the difficulties in the identification of this bacterium using standard microbiological methods. With the introduction of more sensitive and accurate techniques in clinical microbiology, such as genetic approaches and Matrix-Assisted Laser Desorption/Ionization-Time Of Flight (MALDI-TOF) mass spectrometry (MS), the incidence of infections due to A. urinae increased. Herein, we described a case of urinary tract and bloodstream infection caused by A. urinae, which occurred in an 86-year-old Caucasian man with a previous history of prostate cancer. The identification of A. urinae was performed by MALDI-TOF MS, since this microorganism cannot be identified by biochemical reactions. In this report, we highlight the need to consider MALDI-TOF MS as technique of choice for A. urinae identification in the presence of subjects with predisposing factors, such as old age, male gender, and genitourinary tract pathologies.

Differentiation of indoor wood decay fungi with MALDI-TOF mass spectrometry

Holzforschung ◽  
2005 ◽  
Vol 59 (3) ◽  
pp. 374-377 ◽  
Author(s):  
Olaf Schmidt ◽  
Wibke Kallow
Keyword(s):  
Mass Spectrometry ◽  
Wood Decay ◽  
Decay Fungi ◽  
Maldi Tof Ms ◽  
Maldi Tof Mass Spectrometry ◽  
Maldi Tof ◽  
Wood Decay Fungi ◽  
Serpula Lacrymans ◽  
Tof Ms

Abstract MALDI-TOF MS differentiated mycelia within pairs each of the closely related indoor wood decay fungi Serpula lacrymans, S. himantioides, Coniophoraputena, C. marmorata, and Antrodia vaillantii, A. sinuosa. The method is thus suitable to identify unknown samples by spectrum comparison.

scholarly journals Use of Maldi-TOF mass spectrometry in veterinary mycology

2019 ◽  
pp. 20-28
Author(s):  
N. Tyshkivskaya ◽  
A. Tyshkivskaya
Keyword(s):  
Mass Spectrometry ◽  
Animal Feed ◽  
Microscopic Fungi ◽  
Maldi Tof Ms ◽  
Maldi Tof Mass Spectrometry ◽  
Maldi Tof ◽  
Match Rate ◽  
Maldi Biotyper ◽  
Feed Samples ◽  
Tof Ms

Use of MALDI-TOF mass spectrometry to identify yeast and molds in animal feed. The material for the work was animal feed samples received for research from diff erent regions of Ukraine. The presence of yeast and molds was determined according to DSTU ISO 7954:2006. To establish the general contamination of the feed with micromycetes, the fungi were fi rst isolated from the feed by planting them on Saburo medium, and the serial dilution method was used to calculate the content of fungi diaspores in 1 g of feed. The feed samples were incubated and studied at a temperature of 24 ° C for 5–7 days. The identifi cation of molds was carried out using the MALDI-TOF method. In the process of mycological examination of feed during 2018–2019. 198 animal feed samples were examined. During the study period, the largest number of feed was examined, which was 30.4% in 2018, of the total number of samples (19.6% - feed for poultry, 10.8% - for pigs). For fi ve months of 2019, we observed the same trend: in 31.1% of cases, the defi nitions of yeast and molds in compound feeds prevailed, of which 19.8% accounted for compound feeds for poultry and in 11.3% of cases for pigs. In second place in the number of studies, corn samples are 11.9 and 11.3% in 2018 and 2019, respectively. The most common types of fungi in the feed were representatives of the genera Fusarium, Penicillium, Aspergillus, Alternaria, Mucor, Rhizopus, Cladosporium. The affi liation of microscopic fungi to specifi c genera was determined by assessing the morphology of the fungal colony on media and the morphology of conidiophore structures Particular attention was paid to microscopic fungi of the Fusarium family, which are producers of various mycotoxins. Using the MALDI Biotyper software, automatic identifi cation was performed based on a comparison of the collected initial spectra of the fungus with the reference spectra of the database of the instrument itself, as well as with the library of the University of Belgium (BCCM, Belgian Co-Ordinateo collections of micro-organism). Following the results of mass spectrometry, microscopic fungi of the Fusarium family were represented by 9 species. Of these, 5 species were most often found: F. proliferatum, F. acutatum, F. subglutinans, F. verticillioides. Among the fungi of the Aspergillus family, A. fl avus, A. pseudoglaucus, A. tubingensis, and A. niger predominated. Species identifi cation of microscopic fungi using mass spectrometry helps quickly and accurately identify mold fungi and yeast. Determination of the species affi liation of microscopic organisms occurs through analysis of the protein fraction of the lysate of microscopic fungi and yeast ("direct protein profi ling"). MALDI Biotyper software includes automatic identifi cation of molds based on a comparison of the output spectra with the reference spectra of the database. Identifi cation of microorganisms using MALDI-TOF MS is based on the assessment of ribosomal proteins that are usually present in the cell. The sensitivity of the MALDI-TOF MS method is 103106 m.k./cm. In this case, the accuracy of identifi cation depends on the amount of test material. To determine the likelyhood of identifi cation, a given logarithmic indicator is the compliance coeffi cient Score, the value of which is used to evaluate the reliability and adequacy of the results. The higher the match rate, the more likely it is to get the correct identifi cation result. MALDI-TOF technology for mass spectrometric identifi cation of micromycetes has a high measurement speed, low cost of reagents and materials used, and simple preparation holes. MALDI-TOF MS has a high diagnostic sensitivity. Key words: mold identifi cation, MALDI-TOF, mass spectrometry, Fusarium, Penicillium, Aspergillus, Alternaria, Mucor, Rhizopus, Cladosporium.

scholarly journals High-Throughput Quantitative Profiling of Serum N-Glycome by MALDI-TOF Mass Spectrometry and N-Glycomic Fingerprint of Liver Fibrosis

2007 ◽  
Vol 53 (7) ◽  
pp. 1254-1263 ◽  
Author(s):  
Richard KT Kam ◽  
Terence CW Poon ◽  
Henry LY Chan ◽  
Nathalie Wong ◽  
Alex Y Hui ◽  
...  
Keyword(s):  
Mass Spectrometry ◽  
Liver Fibrosis ◽  
High Throughput ◽  
Quantitative Aspect ◽  
Diagnostic Model ◽  
Maldi Tof Ms ◽  
Maldi Tof Mass Spectrometry ◽  
Maldi Tof ◽  
Ishak Score ◽  
Tof Ms

Abstract Background: The use of MALDI-TOF mass spectrometry (MS) in quantitative glycan profiling has not been reported. In this study, we attempted to establish a high-throughput quantitative assay for profiling serum N-glycome, and we applied the new assay to identifying serum N-glycans for diagnosis of liver fibrosis and cirrhosis. Methods: N-glycans from whole serum proteins in 2 μL serum were released by enzymatic digestion, cleaned up by hydrophilic chromatography, and subsequently quantitatively profiled with a linear MALDI-TOF MS system, which was originally designed for quantitative proteomic profiling. Serum N-glycome profiles from 46 patients with chronic hepatitis B infection and with different degrees of liver fibrosis were examined. Results: The intra- and interassay CVs of peak intensities of the standard N-glycans were <8% and <17%, respectively. When the assay was applied to the analysis of serum N-glycome profiles, 17 peaks were found to be potential biomarkers for detection of liver fibrosis/cirrhosis. Linear regression analysis revealed that 4 peaks of 1341.5, 1829.7, 1933.3, and 2130.3 m/z (all P <0.005) had complementary value in detecting liver fibrosis and included them, but not any serological markers, in the diagnostic model. Leave-one-out cross-validation showed the diagnostic model could identify significant fibrosis (Ishak score ≥3) and cirrhosis (Ishak score ≥5), both at 85% accuracy. Conclusion: This is the first study to illustrate the quantitative aspect of MALDI-TOF MS in N-glycome profiling and the first study to reveal the potential value of the serum N-glycan profile for identifying liver fibrosis.

scholarly journals Evaluating Different Storage Media for Identification of Taenia saginata Proglottids Using MALDI-TOF Mass Spectrometry

2021 ◽  
Vol 9 (10) ◽  
pp. 2006
Author(s):  
Tabea P. Wendel ◽  
Maureen Feucherolles ◽  
Jacqueline Rehner ◽  
Sven Poppert ◽  
Jürg Utzinger ◽  
...  
Keyword(s):  
Mass Spectrometry ◽  
Sodium Chloride ◽  
Taenia Saginata ◽  
Maldi Tof Ms ◽  
Maldi Tof Mass Spectrometry ◽  
Maldi Tof ◽  
Storage Media ◽  
Species Specific ◽  
Protein Spectra ◽  
Tof Ms

Taenia saginata is a helminth that can cause taeniasis in humans and cysticercosis in cattle. A species-specific diagnosis and differentiation from related species (e.g., Taenia solium) is crucial for individual patient management and disease control programs. Diagnostic stool microscopy is limited by low sensitivity and does not allow discrimination between T. saginata and T. solium. Molecular diagnostic approaches are not routinely available outside research laboratories. Recently, matrix-assisted laser desorption/ionization time-of-flight (MALDI-TOF) mass spectrometry (MS) was proposed as a potentially suitable technique for species-specific helminth diagnosis. However, standardized protocols and commercial databases for parasite identification are currently unavailable, and pre-analytical factors have not yet been assessed. The purpose of this study was to employ MALDI-TOF MS for the identification of T. saginata proglottids obtained from a human patient, and to assess the effects of different sample storage media on the technique’s diagnostic accuracy. We generated T. saginata-specific main spectral profiles and added them to an in-house database for MALDI-TOF MS-based diagnosis of different helminths. Based on protein spectra, T. saginata proglottids could be successfully differentiated from other helminths, as well as bacteria and fungi. Additionally, we analyzed T. saginata proglottids stored in (i) LC–MS grade water; (ii) 0.45% sodium chloride; (iii) 70% ethanol; and (iv) 37% formalin after 2, 4, 6, 8, 12, and 24 weeks of storage. MALDI-TOF MS correctly identified 97.2–99.7% of samples stored in water, sodium chloride, and ethanol, with log-score values ≥2.5, thus indicating reliable species identification. In contrast, no protein spectra were obtained for samples stored in formalin. We conclude that MALDI-TOF-MS can be successfully employed for the identification of T. saginata, and that water, sodium chloride, and ethanol are equally effective storage solutions for prolonged periods of at least 24 weeks.

scholarly journals DNA-mediated cell surface engineering for multiplexed glycan profiling using MALDI-TOF mass spectrometry

2016 ◽  
Vol 7 (8) ◽  
pp. 5448-5452 ◽  
Author(s):  
Ziyi He ◽  
Qiushui Chen ◽  
Fengming Chen ◽  
Jie Zhang ◽  
Haifang Li ◽  
...  
Keyword(s):  
Mass Spectrometry ◽  
Cell Surface ◽  
Surface Engineering ◽  
Cell Surface Engineering ◽  
Maldi Tof Ms ◽  
Maldi Tof Mass Spectrometry ◽  
Maldi Tof ◽  
Engineering Strategy ◽  
Glycan Profiling ◽  
Tof Ms

A MALDI-TOF MS based approach is developed for multiplexed profiling of cell surface glycans using a DNA-mediated cell surface engineering strategy.

scholarly journals Rapid separation and identification of beer spoilage bacteria by inertial microfluidics and MALDI-TOF mass spectrometry

Lab on a Chip ◽  
2019 ◽  
Vol 19 (11) ◽  
pp. 1961-1970 ◽  
Author(s):  
Mark R. Condina ◽  
Brooke A. Dilmetz ◽  
Sajad Razavi Bazaz ◽  
Jon Meneses ◽  
Majid Ebrahimi Warkiani ◽  
...  
Keyword(s):  
Mass Spectrometry ◽  
High Throughput ◽  
Accurate Method ◽  
Rapid Separation ◽  
Maldi Tof Ms ◽  
Maldi Tof Mass Spectrometry ◽  
Maldi Tof ◽  
Spoilage Bacteria ◽  
Beer Spoilage ◽  
Tof Ms

Microfluidics and MALDI-TOF MS is a rapid, high-throughput, and accurate method for the identification of beer spoilage bacteria.

scholarly journals MALDI-ToF mass spectrometry analysis with molecular genetic identification of Vibrio spp. in the system of the monitoring of vibrio flora of surface water reservoirs

2014 ◽  
Vol 19 (6) ◽  
pp. 27-36
Author(s):  
L. V Mironova ◽  
E. A Basov ◽  
M. V Afanasev ◽  
Zh. Yu Khunkheeva ◽  
S. K Mitkeeva ◽  
...  
Keyword(s):  
Mass Spectrometry ◽  
Surface Water ◽  
Protein Profiling ◽  
Mass Spectrometry Analysis ◽  
Water Reservoirs ◽  
Maldi Tof Ms ◽  
Maldi Tof Mass Spectrometry ◽  
Maldi Tof ◽  
Vibrio Spp ◽  
Tof Ms

There is presented the assessment of the efficacy of the application of direct protein profiling on the basis of MALDI-ToF mass-spectrometry for identification of Vibrio spp. during monitoring of Vibrio flora in surface water reservoirs implemented in the network of cholera surveillance. The comparison of results of the MALDI-ToF MS and bacteriological detection of taxonomic belonging of583 colonies morphologically similar to Vibrio cholerae (isolated in bacteriological examination of samples from environmental objects in Irkutsk city in 2012-2013) with following random identification based on 16S rRNA and rpoB gene structure showed high diagnostic sensitivity and specificity of mass-spectrometry. The findings determine the expediency of the inclusion of MALDI-ToF MS in the layout of microbiological examination in the monitoring of Vibrio flora in surface water reservoirs.

scholarly journals Work Flow Analysis of Around-the-clock Processing of Blood Culture Samples and Integrated MALDI-TOF Mass Spectrometry Analysis for the Diagnosis of Bloodstream Infections

2013 ◽  
Vol 59 (11) ◽  
pp. 1649-1656 ◽  
Author(s):  
Wilhelm Schneiderhan ◽  
Alexander Grundt ◽  
Stefan Wörner ◽  
Peter Findeisen ◽  
Michael Neumaier
Keyword(s):  
Mass Spectrometry ◽  
Blood Culture ◽  
Real Time ◽  
Work Flow ◽  
Pathogen Identification ◽  
Maldi Tof Ms ◽  
Maldi Tof Mass Spectrometry ◽  
Maldi Tof ◽  
Ms Analysis ◽  
Tof Ms

BACKGROUND Because sepsis has a high mortality rate, rapid microbiological diagnosis is required to enable efficient therapy. The effectiveness of MALDI-TOF mass spectrometry (MALDI-TOF MS) analysis in reducing turnaround times (TATs) for blood culture (BC) pathogen identification when available in a 24-h hospital setting has not been determined. METHODS On the basis of data from a total number of 912 positive BCs collected within 140 consecutive days and work flow analyses of laboratory diagnostics, we evaluated different models to assess the TATs for batch-wise and for immediate response (real-time) MALDI-TOF MS pathogen identification of positive BC results during the night shifts. The results were compared to TATs from routine BC processing and biochemical identification performed during regular working hours. RESULTS Continuous BC incubation together with batch-wise MALDI-TOF MS analysis enabled significant reductions of up to 58.7 h in the mean TATs for the reporting of the bacterial species. The TAT of batch-wise MALDI-TOF MS analysis was inferior by a mean of 4.9 h when compared to the model of the immediate work flow under ideal conditions with no constraints in staff availability. CONCLUSIONS Together with continuous cultivation of BC, the 24-h availability of MALDI-TOF MS can reduce the TAT for microbial pathogen identification within a routine clinical laboratory setting. Batch-wise testing of positive BC loses a few hours compared to real-time identification but is still far superior to classical BC processing. Larger prospective studies are required to evaluate the contribution of rapid around-the-clock pathogen identification to medical decision-making for septicemic patients.

scholarly journals Discrimination of Enterobacteriaceae and Non-fermenting Gram Negative Bacilli by MALDI-TOF Mass Spectrometry

2013 ◽  
Vol 7 (1) ◽  
pp. 118-122 ◽  
Author(s):  
Reiner Schaumann ◽  
Nicolas Knoop ◽  
Gelimer H Genzel ◽  
Kevin Losensky ◽  
Christiane Rosenkranz ◽  
...  
Keyword(s):  
Mass Spectrometry ◽  
Salmonella Enterica ◽  
Reference Database ◽  
Alternative Analysis ◽  
Gram Negative ◽  
Maldi Tof Ms ◽  
E Coli ◽  
Maldi Tof Mass Spectrometry ◽  
Maldi Tof ◽  
Tof Ms

Discrimination of Enterobacteriaceae and Non-fermenting Gram Negative Bacilli by MALDI-TOF Mass Spectrometry Matrix assisted laser desorption/ionization time of flight mass spectrometry (MALDI-TOF MS) has proven to be an effective identification tool in medical microbiology. Discrimination to subspecies or serovar level has been found to be challenging using commercially available identification software. By forming our own reference database and using alternative analysis methods, we could reliably identify all implemented Enterobacteriaceae and non-fermenting gram negative bacilli by MALDI-TOF MS and even succeeded to distinguish Shigella sonnei from Escherichia coli (E. coli) and Salmonella enterica spp. enterica serovar Enteritidis from Salmonella enterica spp. enterica serovar Typhimurium. Furthermore, the method showed the ability to separate Enterohemorrhagic E. coli (EHEC) and Enteropathogenic E. coli (EPEC) from non-enteropathogenic E. coli.

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