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      Performance of MALDI biotyper compared with Vitek ™ 2 compact system for fast identification and discrimination of Staphylococcus species isolated from bovine mastitis

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          Abstract

          This study was designed to evaluate the ability of MALDI Biotyper ( MBT) compared with Vitek ™ 2 compact system for accurate identification of Staphylococcus aureus ( S. aureus) and coagulase‐negative staphylococci ( CNS) strains and discriminate methicillin‐sensitive S. aureus ( MSSA) from methicillin‐resistant S. aureus ( MRSA). Throughout Al‐Qassim region, Saudi Arabia, a total of 198 isolates of S. aureus (132 MSSA and 66 MRSA) and 44 CNS were collected from five dairy farms where the prevalence of staphylococcal mastitis was reported. The results produced by Vitek ™ 2 compact system demonstrated that 123/132 MSSA isolates (93.18%), 61/66 MRSA (92.42%), and 37/44 CNS species (84.09%) were correctly identified. However; 130/132 MSSA (98.48%), 64/66 MRSA (96.96%), and 44/44 CNS (100%) were correctly identified by MBT with score ≥2. 00. The principal component analysis ( PCA) dendrogram generated by MBT illustrated that the tested isolates were classified into two groups of Staphylococcus species at the distance level of 600. S. aureus isolates were found to be closely related with higher peak intensities in the mass of 3,993 Da, 4,121 Da and 5,845 Da were detected in MRSA, whereas, that were lost in MSSA. Conclusion: This study verified that MBT is an alternative powerful tool for precise identification and discrimination of Staphylococcus species.

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          Ongoing revolution in bacteriology: routine identification of bacteria by matrix-assisted laser desorption ionization time-of-flight mass spectrometry.

          Piseth Seng, Michel Drancourt, Frédérique Gouriet … (2009)
          Matrix-assisted laser desorption ionization time-of-flight (MALDI-TOF) mass spectrometry accurately identifies both selected bacteria and bacteria in select clinical situations. It has not been evaluated for routine use in the clinic. We prospectively analyzed routine MALDI-TOF mass spectrometry identification in parallel with conventional phenotypic identification of bacteria regardless of phylum or source of isolation. Discrepancies were resolved by 16S ribosomal RNA and rpoB gene sequence-based molecular identification. Colonies (4 spots per isolate directly deposited on the MALDI-TOF plate) were analyzed using an Autoflex II Bruker Daltonik mass spectrometer. Peptidic spectra were compared with the Bruker BioTyper database, version 2.0, and the identification score was noted. Delays and costs of identification were measured. Of 1660 bacterial isolates analyzed, 95.4% were correctly identified by MALDI-TOF mass spectrometry; 84.1% were identified at the species level, and 11.3% were identified at the genus level. In most cases, absence of identification (2.8% of isolates) and erroneous identification (1.7% of isolates) were due to improper database entries. Accurate MALDI-TOF mass spectrometry identification was significantly correlated with having 10 reference spectra in the database (P=.01). The mean time required for MALDI-TOF mass spectrometry identification of 1 isolate was 6 minutes for an estimated 22%-32% cost of current methods of identification. MALDI-TOF mass spectrometry is a cost-effective, accurate method for routine identification of bacterial isolates in or =10 reference spectra per bacterial species and a 1.9 identification score (Brucker system). It may replace Gram staining and biochemical identification in the near future.
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            Performance of matrix-assisted laser desorption ionization-time of flight mass spectrometry for identification of bacterial strains routinely isolated in a clinical microbiology laboratory.

            G Prod'hom, A Bizzini, G Greub … (2010)
            Matrix-assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF MS) has recently been introduced in diagnostic microbiology laboratories for the identification of bacterial and yeast strains isolated from clinical samples. In the present study, we prospectively compared MALDI-TOF MS to the conventional phenotypic method for the identification of routine isolates. Colonies were analyzed by MALDI-TOF MS either by direct deposition on the target plate or after a formic acid-acetonitrile extraction step if no valid result was initially obtained. Among 1,371 isolates identified by conventional methods, 1,278 (93.2%) were putatively identified to the species level by MALDI-TOF MS and 73 (5.3%) were identified to the genus level, but no reliable identification was obtained for 20 (1.5%). Among the 1,278 isolates identified to the species level by MALDI-TOF MS, 63 (4.9%) discordant results were initially identified. Most discordant results (42/63) were due to systematic database-related taxonomical differences, 14 were explained by poor discrimination of the MALDI-TOF MS spectra obtained, and 7 were due to errors in the initial conventional identification. An extraction step was required to obtain a valid MALDI-TOF MS identification for 25.6% of the 1,278 valid isolates. In conclusion, our results show that MALDI-TOF MS is a fast and reliable technique which has the potential to replace conventional phenotypic identification for most bacterial strains routinely isolated in clinical microbiology laboratories.
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              Comparison of two matrix-assisted laser desorption ionization-time of flight mass spectrometry methods with conventional phenotypic identification for routine identification of bacteria to the species level.

              Jacques Schrenzel, Abdessalam Cherkaoui, Stéphane Emonet … (2010)
              Bacterial identification relies primarily on culture-based methodologies requiring 24 h for isolation and an additional 24 to 48 h for species identification. Matrix-assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF MS) is an emerging technology newly applied to the problem of bacterial species identification. We evaluated two MALDI-TOF MS systems with 720 consecutively isolated bacterial colonies under routine clinical laboratory conditions. Isolates were analyzed in parallel on both devices, using the manufacturers' default recommendations. We compared MS with conventional biochemical test system identifications. Discordant results were resolved with "gold standard" 16S rRNA gene sequencing. The first MS system (Bruker) gave high-confidence identifications for 680 isolates, of which 674 (99.1%) were correct; the second MS system (Shimadzu) gave high-confidence identifications for 639 isolates, of which 635 (99.4%) were correct. Had MS been used for initial testing and biochemical identification used only in the absence of high-confidence MS identifications, the laboratory would have saved approximately US$5 per isolate in marginal costs and reduced average turnaround time by more than an 8-h shift, with no loss in accuracy. Our data suggest that implementation of MS as a first test strategy for one-step species identification would improve timeliness and reduce isolate identification costs in clinical bacteriology laboratories now.
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                Author and article information

                Contributors
                Ayman Elbehiry: ar.elbehiry@qu.edu.sa
                Journal
                Journal ID (nlm-ta): Microbiologyopen
                Journal ID (iso-abbrev): Microbiologyopen
                Journal ID (doi): 10.1002/(ISSN)2045-8827
                Journal ID (publisher-id): MBO3
                Title: MicrobiologyOpen
                Publisher: John Wiley and Sons Inc. (Hoboken )
                ISSN (Electronic): 2045-8827
                Publication date (Electronic): 01 July 2016
                Publication date Collection: December 2016
                Volume: 5
                Issue: 6 ( doiID: 10.1002/mbo3.2016.5.issue-6 )
                Pages: 1061-1070
                Affiliations
                [ 1 ] Department of Bacteriology Mycology and Immunology Faculty of Veterinary MedicineSadat City University SadatEgypt
                [ 2 ] Department of Public Health Microbiology Unit College of Public Health and Health InformaticsQassim University BuraidahSaudi Arabia
                [ 3 ] Department of Veterinary Medicine College of Agriculture and Veterinary MedicineQassim University BuraidahSaudi Arabia
                [ 4 ] Department of Medical laboratories College of Applied Medical ScienceQassim University BuraidahSaudi Arabia
                [ 5 ] Department of Animal Medicine Faculty of Veterinary MedicineKafrelsheikh University BuraidahEgypt
                [ 6 ] Department of Physiology Faculty of MedicineZagazig University ZagazigEgypt
                Author notes
                [*] [* ] Correspondence

                Ayman Elbehiry, Department of Public Health College of Public Health and Health Informatics University of Qassim PO. Box 52571, Buraydah, Saudi Arabia

                Tel: +966 505 72 17, Fax: +9716 558 58 79, Mobile: +966532207969

                Email: ar.elbehiry@ 123456qu.edu.sa

                Alternative e‐mail: aymanella2007@ 123456yahoo.com

                Article
                Publisher ID: MBO3389
                DOI: 10.1002/mbo3.389
                PMC ID: 5221440
                PubMed ID: 27364641
                SO-VID: bf55c61f-b603-47f5-b9ff-e53a6a7b60bd
                Copyright © © 2016 The Authors. MicrobiologyOpen published by John Wiley & Sons Ltd.
                License:

                This is an open access article under the terms of the Creative Commons Attribution License, which permits use, distribution and reproduction in any medium, provided the original work is properly cited.

                History
                Date received : 18 April 2016
                Date revision received : 23 May 2016
                Date accepted : 28 May 2016
                Page count
                Figures: 5, Tables: 3, Pages: 10, Words: 7079
                Funding
                Funded by: National Science, Technology and Innovation Plan (NSTIP)
                Award ID: 13‐BIO1683‐09
                Categories
                Subject: Original Research
                Subject: Original Research
                Custom metadata
                source-schema-version-number 2.0
                component-id mbo3389
                cover-date December 2016
                details-of-publishers-convertor Converter:WILEY_ML3GV2_TO_NLMPMC version:5.0.0 mode:remove_FC converted:09.01.2017

                ScienceOpen disciplines: Microbiology & Virology
                Keywords: discrimination,identification,maldi biotyper,mrsa,mssa,staphylococcus species
                Data availability:
                ScienceOpen disciplines: Microbiology & Virology
                Keywords: discrimination, identification, maldi biotyper, mrsa, mssa, staphylococcus species

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