Microflex maldi biotyper

Manufactured by Bruker

Sourced in Germany, United States

The Microflex MALDI Biotyper is a compact and versatile laboratory instrument designed for rapid and accurate microbial identification. It utilizes matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) technology to generate unique protein fingerprints from bacterial and fungal samples.

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Spelling variants of this product

MALDI Biotyper (289 citations) Microflex (153 citations) MALDI Biotyper Microflex LT (15 citations) Biotyper Microflex (10 citations) MALDI Biotyper Microflex system (4 citations) Microflex LT-MALDI Biotyper mass spectrometer (4 citations) Microflex LT-MALDI Biotyper System (4 citations) Maldi BioTyper microflex LT/SH (3 citations) MALDI Microflex LT and Biotyper database (2 citations)

23 protocols using microflex maldi biotyper

Bacterial Strain Identification and Antibiotic Susceptibility

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In the enrolled samples, we noted replicate number identified, mono or polymicrobial infection with associated baterium (bacterial species) and identification scores of bacterial strains. The strains were identified directly after inoculation of samples on the appropriate agar medium, except the dialysis fluid, which was detected negative after incubation. The strain was identified after incubation on Schaedler broth + 0.02% agar (+ Vit. K3) (BioMérieux, Lyon, France) and on aerobic and anaerobic blood culture bottles (Bact/ALERT^® FA Plus (aerobic) and FN Plus (anaerobic) bottles, BioMérieux, Lyon, France). Liquid mediums were then inoculated on the standard agar plate. L. adecarboxylata was identified using MALDI-TOF MS with an identification score of 2.0 (Table 1). For all samples, direct detection and identification (from agar medium, without extraction) used MALDI-TOF MS (MALDI Biotyper-Microflex^®, Bruker Daltonics, Bremen, Germany) with Tryptic soy agar with 5% sheep blood, except urine with UriSelect 4 Medium, Bio-Rad, a non-selective chromogenic agar medium.
Antibiotic susceptibility was tested by the disc diffusion assay (disc and Mueller Hinton agar: Biorad, Marnes-la-Coquette, France). CASFM/EUCAST breakpoints were used for the interpretation (Table 2).

Zayet S., Lang S., Garnier P., Pierron A., Plantin J., Toko L., Royer P.Y., Villemain M., Klopfenstein T, & Gendrin V. (2021). Leclercia adecarboxylata as Emerging Pathogen in Human Infections: Clinical Features and Antimicrobial Susceptibility Testing. Pathogens, 10(11), 1399.

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Bacterial Strains Characterization and Preparation

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Pseudomonas aeruginosa PAO1 wild type was obtained from the Institute Pasteur Collection (CIP 104116, Paris, France). Methicillin-resistant Staphylococcus aureus (MRSA, ATCC® 33591™) and Burkholderia cepacia (ATCC® 25416™) were both obtained from the ATCC® (Manassas, USA). Clinical strains: one mucoid P. aeruginosa and two S. aureus strains: one methicillin-sensitive (MSSA) and one methicillin-resistant (MRSA), were from the Collection of Purpan hospital (Toulouse, France), isolated from CF patients on cetrimide and mannitol salt agar, respectively (Table 2). Identification was performed regarding macroscopic and microscopic characteristics and biochemical properties (oxidase, catalase). Finally, strains were identified at the species level by MALDI-TOF MS (Maldi Biotyper Microflex®, Bruker Daltonics, Bremen, Germany; IVD 7712) (score ≥2.2). In the present study, the bacterial strains will be referred to as follows:
Strains were frozen and kept at −80 °C in a 20% (v/v) glycerol stock solution. Before each experiment, two successive subcultures were prepared on trypticase soy agar (TSA, Biomérieux, Craponne, France) and incubated for 24 h under aerobic conditions at 37 °C. Before each experiment, 10⁸ CFU/mL bacterial suspension was prepared by adjusting OD at 640 nm to 0.150.

Trognon J., Rima M., Lajoie B., Roques C, & El Garah F. (2023). NaCl-induced modulation of species distribution in a mixed P. aeruginosa / S. aureus /B.cepacia biofilm. Biofilm, 6, 100153.

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Comprehensive Identification and Antibiotic Resistance Profiling of Bacterial Isolates

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Strain identification was based on Gram staining, API 20 Strep system (bioMérieux, Marcy l’Etoile, France), and matrix-assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF MS) (MALDI Biotyper Microflex™; Bruker Daltonics, Bremen, Germany), and confirmed by specific amplification of ddI genes encoding the d-Ala-d-Ala ligase as previously described.16 (link) Antibiotic susceptibility testing was performed by using the disk diffusion method. For glycopeptides, minimum inhibitory concentrations (MICs) of vancomycin and teicoplanin were determined by the E-test method (bioMérieux). Results were interpreted according to the recommendations of the Clinical and Laboratory Standards Institute (CLSI) and MDR defined according to Magiorakos et al.17 ,18 (link) Multiplex PCR assay allowing simultaneous and specific detection of vanA, vanB, vanC-1, and vanC-2/3 resistance genes was performed according to the method previously described with no previous DNA extraction and no further sequencing required.16 (link)

Benammar S., Pantel A., Aujoulat F., Benmehidi M., Courcol R., Lavigne J.P., Romano-Bertrand S, & Marchandin H. (2018). First molecular characterization of related cases of healthcare-associated infections involving multidrug-resistant Enterococcus faecium vanA in Algeria. Infection and Drug Resistance, 11, 1483-1490.

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Species Identification via MALDI-TOF MS

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The verification of species identification was performed via matrix-assisted laser desorption ionization–time of flight (MALDI-TOF) mass spectrometry (MALDI Biotyper, Microflex, Bruker Daltonics GmbH, Bremen, Germany).

Wohlfarth E., Kresken M., Deuchert F., Gatermann S.G., Pfeifer Y., Pfennigwerth N., Seifert H., Higgins P.G, & Werner G. (2023). In Vitro Activity of Cefiderocol against Clinical Gram-Negative Isolates Originating from Germany in 2016/17. Antibiotics, 12(5), 864.

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MALDI-TOF MS for Enterococcus Identification

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Identification of isolated Enterococcus spp. strains was performed using the MALDI BioTyper Microflex mass spectrometer (Bruker, Billerica, MA, USA), which uses the MALDI-TOF MS (Matrix Assisted Laser Desorption Laser–Time of Flight Mass Spectrometry), following the manufacturer’s recommendations. The probability of assigning a microorganism to an appropriate species was determined on the basis of the parameter described in the system as score value, the value of which was assessed in accordance with the manufacturer’s instructions (score value > 2.000 means the highly probable assignment of the microorganism to the species).

Kraszewska Z., Skowron K., Kwiecińska-Piróg J., Grudlewska-Buda K., Przekwas J., Wiktorczyk-Kapischke N., Wałecka-Zacharska E, & Gospodarek-Komkowska E. (2022). Antibiotic Resistance of Enterococcus spp. Isolated from the Urine of Patients Hospitalized in the University Hospital in North-Central Poland, 2016–2021. Antibiotics, 11(12), 1749.

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MALDI-Biotyper Strain Identification

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About 5–10 mg of cells was suspended in 300 μl of sterile distilled water in a micro centrifuge tube. A series of three washes were given with 900 μl absolute ethanol and 50 μl of 70% formic acid followed by centrifugation at 12000 rpm for 2 min. The pellet was dissolved in 50 μL acetonitrile and centrifuged at 12000 rpm for 2 min. One microliter from the supernatant was loaded on target and dried followed by analysis using MALDI-Biotyper Microflex, Bruker, Germany. The MALDI-biotyping for strain identification was conducted at the MTCC, Council of Scientific and Industrial Research-Institute of Microbial Technology (CSIR-IMTECH), India.

Chalasani A.G., Dhanarajan G., Nema S., Sen R, & Roy U. (2015). An Antimicrobial Metabolite from Bacillus sp.: Significant Activity Against Pathogenic Bacteria Including Multidrug-Resistant Clinical Strains. Frontiers in Microbiology, 6, 1335.

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MALDI-ToF MS Species Confirmation

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One putative colony of CP E. coli of each agar plate (six per sample) was chosen for species confirmation by MALDI-ToF MS. As matrix α-Cyano-4-hydroxycinnamic acid (HCCA, Bruker, USA) was used, and analyses were performed by MALDI Microflex Biotyper (Bruker Daltonics, Bremen, Germany) as recommended by the manufacturers.

Pauly N., Klaar Y., Skladnikiewicz-Ziemer T., Juraschek K., Grobbel M., Hammerl J.A., Hemmers L., Käsbohrer A., Schwarz S., Meemken D., Tenhagen B.A, & Irrgang A. (2021). Isolation Procedure for CP E. coli from Caeca Samples under Review towards an Increased Sensitivity. Microorganisms, 9(5), 1105.

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MALDI-TOF Profiling of Bacterial Strains

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MALDI–TOF mass spectrometry profiles of the strains were obtained using a MALDI Microflex Biotyper (Bruker, Billerica, MA, USA) with parameters as follows: mass range 1960–20,137; ion source 1: 20 kV, 2: 18 kV; lens: 6 kV; detector gain voltage: 2500 V (linear base). For each strain, a cultured colony on the agar plate was taken by a wooden toothpick and smeared onto a target plate (MSP 96 target polished steel BC; Bruker) and fixed with 1 μL HCCA (α-cyano-4-hydroxycinnamic acid) (Bruker). Two hundred laser shots were shot at one point of the target to acquire an accumulated spectrum of 200 spectra, and if the peak height reached a preset threshold, the spectrum was retained. The point in the target was randomly moved. When six accumulated spectra were obtained, all six were added and finalized. The spectra were obtained in triplicate at least, and the representative spectrum with the clearest and rich pattern was selected. The raw spectral data were analyzed using flexAnalysis v3.4 software (Bruker). The spectra data were submitted to jPOST Repository under accession number JPST001137 (PXID: PXD025627) (https://repository.jpostdb.org/entry/JPST001137, accessed on 7 June 2021).

Dekio I., Sugiura Y., Hamada-Tsutsumi S., Murakami Y., Tamura H, & Suematsu M. (2021). What Do We See in Spectra?: Assignment of High-Intensity Peaks of Cutibacterium and Staphylococcus Spectra of MALDI-TOF Mass Spectrometry by Interspecies Comparative Proteogenomics. Microorganisms, 9(6), 1243.

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Antimicrobial Susceptibility of Group B Streptococcus

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Each swab specimen was inoculated on a 5% blood agar plate (Synergy Innovation, Seongnam, Korea) and incubated at 35℃–37℃ in a 5% CO₂ incubator for 24 h. For GBS detection, suspicious colonies showing large, gray, and translucent colonies with a narrow zone of beta-hemolysis or non-beta-hemolysis on blood agar plates were identified using a Microflex MALDI Biotyper (Bruker Daltonik GmbH, Bremen, Germany). A total of 3512 GBS isolates was tested for antimicrobial susceptibility to ampicillin, penicillin, cefotaxime, ceftriaxone, cefepime, meropenem, levofloxacin, erythromycin, clindamycin, tetracycline, chloramphenicol, and vancomycin using the Vitek 2 automated system (bioMérieux, Marcy l’Etoile, France).15

Bae H.G., Hong J., Kim Y.J., Lee K.R., Lee K., Choi S.J, & Uh Y. (2022). A Retrospective National Study on Colonization Rate and Antimicrobial Susceptibility of Streptococcus agalactiae in Pregnant Korean Women, 2018–2020. Yonsei Medical Journal, 63(8), 717-723.

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Urine Culture and Bacterial Identification

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10 µL of each un-centrifuged urine sample was cultured on UriSelect chromogenic agar plates (Bio-Rad, Berkeley, CA, USA) with a calibrated loop, according to the manufacturer’s instructions and incubated at 37 °C for 24–48 hours, aerobically. If the relevant pathogens presented in significant colony count, the plates were passed on for further processing. Between 2008–2012, presumptive phenotypic (biochemical reaction-based) methods and VITEK 2 Compact Automated ID/AST System (bioMérieux, Marcy-l'Étoile, France) were used for bacterial identification, and, after 2013, this was complemented by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS; Bruker Daltonik Gmbh. Gr.). The methodology of sample preparation for MALDI-TOF MS measurements was described elsewhere [61 (link)]. Mass spectrometry was performed by the Microflex MALDI Biotyper (Bruker Daltonics, Germany) in positive linear mode across the m/z range of 2 to 20 kDa; for each spectrum, 240 laser shots at 60 Hz in groups of 40 shots per sampling area were collected. The MALDI Biotyper RTC 3.1 software (Bruker Daltonics, Germany) and the MALDI Biotyper Library 3.1 were used for spectrum analysis.

Gajdács M, & Urbán E. (2019). Comparative Epidemiology and Resistance Trends of Proteae in Urinary Tract Infections of Inpatients and Outpatients: A 10-Year Retrospective Study. Antibiotics, 8(3), 91.

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