The swabs were placed into tubes containing 5 mL of Brain Heart Infusion (BHI) broth (LiofilChem, Via Scozia, Livorno, Italy) supplemented with 6.5% NaCl and incubated at 37 °C for 24 h. Subsequently, the inoculum was applied to CHROMagar MRSA (CHROMagar, Paris, France) plates to facilitate the isolation of MRSA. Up to three colonies displaying S. aureus characteristics but exhibiting morphological variations were collected from each plate. Identification of the S. aureus species was conducted through biochemical tests (catalase, DNase, and coagulase) and Bruker Biotyper MALDI-TOF MS (Bruker Daltonics, Bremen, Germany).
Maldi tof ms biotyper
Manufactured by Bruker
Sourced in Germany, United States
The MALDI-TOF MS Biotyper is a mass spectrometry-based identification system for the rapid and reliable identification of microorganisms, including bacteria and yeasts. It utilizes matrix-assisted laser desorption/ionization time-of-flight (MALDI-TOF) technology to generate unique protein fingerprints for the identification of microorganisms.
Automatically generated - may contain errors
Lab products found in correlation
Bhi broth, by Liofilchem (2 mentions)
Uriselect agar, by Bio-Rad (2 mentions)
Microflex lt, by Bruker (2 mentions)
Flexcontrol 3, by Bruker (2 mentions)
Drigalski agar plates, by bioMérieux (1 mentions)
Biotyper, by Bruker (1 mentions)
Abi 3730xl system, by Thermo Fisher Scientific (1 mentions)
Amoxicillin, by bioMérieux (1 mentions)
Oxacillin, by BD (1 mentions)
Pbp2a sa culture colony test, by Abbott (1 mentions)
Amoxicillin clavulanic acid, by bioMérieux (1 mentions)
Oxacillin, by bioMérieux (1 mentions)
Macconkey agar, by BD (1 mentions)
Msupercarba, by CHROMagar (1 mentions)
Buffered peptone water (bpw), by Thermo Fisher Scientific (1 mentions)
Macconkey, by Thermo Fisher Scientific (1 mentions)
Fluconazole, by Thermo Fisher Scientific (1 mentions)
Chloramphenicol, by Thermo Fisher Scientific (1 mentions)
Microbank vial, by Pro-Lab Diagnostics (1 mentions)
Sterile buffered peptone water, by Thermo Fisher Scientific (1 mentions)
38 protocols using maldi tof ms biotyper
1
Isolation and Identification of MRSA
2
Identifying Environmental Source of Enterobacter Outbreak
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In January 2017, the LICT set up an environment sampling campaign to identify a possible environmental source of the outbreak. Multiple environmental sites were tested, including shared devices in the ward (gloves, sheets, plaster, ultrasound gel, neonatal incubators, etc.) using contact plates or swabs. In addition, water samples were collected in different rooms and filtered to search for Enterobacter spp., and multiple siphons were swabbed. The swabs were inoculated on Columbia agar with 5.0% sheep blood and Drigalski agar plates (bioMérieux SA, Marcy l’Etoile, France). The isolated colonies were identified using the reference spectra library of the Bruker Biotyper MALDI-TOF MS (Bruker Daltonics). After the absence of identification of the contamination source and as the outbreak was still active, in July 2017, we implemented two different incubator sampling protocols. Both methodologies included the sampling of the corners and of risky and unattainable areas (seals, ventilator, holes, etc.) just after cleaning. The first method was performed under “off” conditions, and the second was performed under “on” conditions, which were 37°C, and 85.0% humidity for 48 h. Both methods were used on the two incubator models (model A, n = 22; and model B, n = 11) owned by the NICU.
3
Characterization of C. striatum Isolates
4
MALDI-TOF MS for Microbial Identification
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Bruker Biotyper MALDI-TOF MS was implemented in October 2013. It replaced identification of all isolates by the conventional methods listed in the above paragraph. Times to results for positive isolates, preliminary negative results, and antimicrobial testing reports were as described above.
5
Microbial Identification using MALDI-TOF MS
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Identification of isolates was performed in the Houston Methodist clinical microbiology laboratory using a Bruker Biotyper MALDI-TOF MS as part of the standard clinical microbiology practice as described previously (19 (link)). Briefly, colonial material from agar plates or pelleted cells from liquid blood cultures were transferred to the target plate, dried at room temperature for approximately 2 min, and covered with alpha-cyano-4-hydroxycinnamic acid (HCCA) matrix. Spectra were collected using the default instrument settings and interpreted using the research use only microorganism reference library on the Biotyper (Bruker, Billerica, MA). At the time of isolation in 2011 to 2015, Bruker microorganism reference library versions starting with 4.0.0.0 were used. The strains were reanalyzed in 2017 using version 6.0.0.0, which was installed in January 2017.
6
Identification of Cryptococcus Isolates
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Amplification of the rDNA internal transcribed spacer (ITS) region was performed as previously described.15 (link), 19 (link) The PCR products were sequenced in both directions using the DNA analyzer ABI 3730XL system (Applied Biosystems, Foster City, CA). The obtained ITS sequences of Cryptococcus isolates were compared against those contained in the Centraalbureau voor Schimmelcultures (CBS) Fungal Biodiversity Centre database by using BIOLOMICSNET software (http://www.cbs.knaw.nl/collections/BioloMICSSequences.aspx) .
Bruker Biotyper MALDI-TOF MS (Bruker, Daltonik, Bremen, Germany) was also used to identify the conserved 98 Cryptococcus clinical strains. According to the manufacture’s recommendations and as Buchan et al. described, 16 identification by plate-grown isolates (on-plate protein extraction method) was applied. The procedure was performed as previously reported.15 (link)
Bruker Biotyper MALDI-TOF MS (Bruker, Daltonik, Bremen, Germany) was also used to identify the conserved 98 Cryptococcus clinical strains. According to the manufacture’s recommendations and as Buchan et al. described, 16 identification by plate-grown isolates (on-plate protein extraction method) was applied. The procedure was performed as previously reported.15 (link)
7
CRKP Surveillance at Chinese Hospital
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A laboratory-based surveillance of CRKP was conducted between July 2017 and June 2018 at Children’s Hospital affiliated to Zhengzhou University, a 2200-bed tertiary-care teaching hospital located in Henan Province, China, with over 95,000 inpatients per year. The neonatal intensive care unit (NICU I), general neonatal wards (NICU II) and premature wards together comprise 200 beds. The strains were isolated from specimens of sputum, urine, blood, alveolar lavage and cerebrospinal fluid. All suspected isolates were confirmed as K. pneumoniae using a Bruker Biotyper MALDI-TOF MS (Bruker Daltonik GmbH, Bremen, Germany). Among these isolates, CRKP isolates were identified as resistant to Imipenem (IMP) or Meropenem (MEM) by broth microdilution method. Escherichia coli ATCC 25,922 was used as a quality control.19
The study protocol complied with the Declaration of Helsinki and was approved by the Children’s Hospital affiliated to Zhengzhou University Ethics Committee. Informed patient consent was not required as this study had no impact on the patients, and patient information was anonymized.
The study protocol complied with the Declaration of Helsinki and was approved by the Children’s Hospital affiliated to Zhengzhou University Ethics Committee. Informed patient consent was not required as this study had no impact on the patients, and patient information was anonymized.
8
Microbial Identification and MRSA Characterization
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Microbial identification was confirmed by Bruker Biotyper MALDI-TOF MS (Bruker, Billerica, MA). For MRSA designation, a PBP2a SA culture colony test (Alere) was performed according to the manufacturer’s instructions using 18- to 24-h subculture growth. The presence of mecA and the homolog mecC were determined by in-house PCRs (22 (link), 23 (link)). Susceptibility testing included that for cefoxitin by disk diffusion and for oxacillin by disk diffusion and gradient diffusion. Methods followed the procedural guidelines outlined by the Clinical and Laboratory Standards Institute (documents M02 and M23) (55 , 56 ). Disk diffusion testing of cefoxitin (Hardy Diagnostics) and oxacillin (BD) was performed on conventional Mueller-Hinton agar (MHA) (Hardy Diagnostics). Gradient diffusion testing of oxacillin (bioMérieux) was performed on MHA with 2% NaCl agar (Remel). Detection of beta-lactamase production was assessed by the disk diffusion penicillin zone edge test (Hardy Diagnostics) and nitrocefin-based Cefinase disk test (Hardy Diagnostics). Beta-lactamase inhibitor rescue phenotype was determined by assessing the fold change in MIC from amoxicillin (bioMérieux) and amoxicillin-clavulanic acid (bioMérieux) gradient diffusion testing.
9
Carbapenem-resistant Enterobacteriaceae Detection
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Rectal swabs were inoculated onto MacConkey agar (BD, USA) supplemented with ceftriaxone (4 mg/L) and the plates incubated at 37 °C for 18 h. Bacterial identification was performed using Biotyper MALDI-TOF MS (Bruker Daltonics, Germany) according to the manufacturer’s protocol. Colonies identified as Enterobacteriaceae were tested for antimicrobial susceptibility using standard methods and following the guidelines for the disk-diffusion method [15 ]. Confirmation of suspected carbapenemase production in Enterobacteriaceae-positive specimens was performed using a modified carbapenem inactivation method [16 ]. Phenotypic screening for the presence of carbapenemases was performed using a double-disc synergy approach with phenylboronic acid or ethylenediaminetetraacetic acid with meropenem as previously described [17 (link)]. Colistin resistance was tested using the broth microdilution method with cation-adjusted Mueller–Hinton II broth [16 ]. Susceptibility to tigecycline was not tested at Siriraj Hospital during the study period.
10
Isolation and Identification of Multidrug-Resistant Bacteria
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From each environmental sample 25 g material was collected and then diluted 1:10 in Buffered Pepton Water (BPW). The 25 g material and BPW was mixed with a spoon and incubated at 37°C for 18–20 h as shown in Figure 1 . One ml of the pre-enrichment was then diluted 1:10 with Peptone salt water, and from this dilution 10 μl was transferred to a CHROMagar™ C3GR, CHROMagar™ mSuperCARBA™ and CHROMagar™ COL-APSE plate, respectively, and the agar plates were incubated at 37°C overnight. Suspected colonies were identified based on the colony morphology as described by the manufacturer, and sub-streaked on the same selective agar again. Bacterial species identification was performed using a Bruker Biotyper MALDI-TOF MS.
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