The Klebsiella Pneumoniae isolates were plated on Columbia blood agar (bioMérieux, Marcy l'Étoile, France) and incubated for 18 h to 24 h at 37°C. Isolated colonies of each strain were selected and used for MALDI-TOF MS identification using the MALDI-TOF MS (BioMerieux, Marcy-l'Etoile, France), as previously described (Rodel et al., 2019 (link)). The obtained spectra were manually selected in the spectra mode of SARAMIS Premium software (BioMerieux, Marcy-l'Etoile, France). Cluster analysis were performed by spectra compared to each other in SARAMIS RUO database according to the manufacturer's instructions (Vitek MS Plus SARAMIS Premium user manual, BioMerieux, Marcy-l'Etoile, France). Consensus spectra were analyzed with a single link agglomerative clustering algorithm, applying the relative taxonomy analysis tool of SARAMIS Premium software to show the resulting dendrogram with differences and similarities in relative terms (percent matching masses). As a standard setting, the mass signal intensity was not considered in the cluster analysis. According to the type assignment, we defined a cut-off value was >75% similarity (Meng et al., 2019 (link)).
Maldi tof ms
Manufactured by bioMérieux
Sourced in France, United States, Germany
The MALDI-TOF MS is a laboratory instrument used for the identification and characterization of microorganisms. It utilizes Matrix-Assisted Laser Desorption/Ionization (MALDI) and Time-of-Flight (TOF) mass spectrometry techniques to rapidly analyze microbial samples.
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Lab products found in correlation
Vitek 2 system, by bioMérieux (14 mentions)
Vitek 2 compact system, by bioMérieux (13 mentions)
Vitek 2, by bioMérieux (13 mentions)
Vitek 2 compact, by bioMérieux (12 mentions)
Vitek ms, by bioMérieux (9 mentions)
Vitek 2 compact ast gn16, by bioMérieux (4 mentions)
Luria bertani broth medium, by Thermo Fisher Scientific (4 mentions)
Vitek 2 automated system, by bioMérieux (3 mentions)
Vitek 2 gn09, by bioMérieux (2 mentions)
Sabouraud dextrose agar, by Thermo Fisher Scientific (2 mentions)
Vitek 2 compact ast gn67, by bioMérieux (2 mentions)
Bactec plus aerobic f, by BD (2 mentions)
Vitek2 gp card, by bioMérieux (2 mentions)
Mueller hinton agar plate, by Thermo Fisher Scientific (2 mentions)
Fossmatic system 4000, by Foss (2 mentions)
Columbia sheep blood agar, by Thermo Fisher Scientific (2 mentions)
Luria bertani broth medium, by Merck Group (2 mentions)
Etest, by bioMérieux (2 mentions)
Eugenol, by MedChemExpress (2 mentions)
Thymol, by Merck Group (2 mentions)
184 protocols using maldi tof ms
1
MALDI-TOF MS Identification of Klebsiella Pneumoniae
2
Genetic Characterization of ESBL-Producing Klebsiella pneumoniae
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Bacteria were identified using MALDI-TOF MS (bioMérieux; Marcy l’Étoile, France), and antibiotic susceptibility tests were performed by the broth microdilution method using the Vitek-2 system (bioMérieux). Isolates of KPN showing ESBL resistance were analyzed by PFGE and multi-locus sequence typing to determine their genetic relatedness. Once the isolates were digested with XbaI (Roche, Basel, Switzerland) enzyme, electrophoresis was performed using CHEF MAPPER (Bio-Rad, Hercules, CA, USA). The agarose gel was stained with SYBR Gold (ThermoFisher Scientific, Waltham, MA, USA) to visualize the PFGE pattern. A dendrogram was obtained using BioNumerics (Bio-Rad) to evaluate the relationship between the strains. The similarity in PFGE patterns was interpreted according to the criteria of Tenover et al. [15 (link)]. Seven housekeeping genes (rpoB, gapA, mdh, pgi, phoE, infB, and tonB) were amplified and sequenced to identify STs as described at http://bigsdb.pasteur.fr/klebsiella/ [16 (link)]. The ESBL gene was amplified by PCR with known primers targeting the CTX-M-1, CTX-M-2, and CTX-M-9 genes [17 (link), 18 (link)]. DNA sequencing was performed using the amplified PCR product, and ESBL genotypes were identified using BLAST.
3
Antibiotic Resistance Profiling
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The VITEK 2 system and the MALDI TOF MS (bioMérieux, France) were applied for isolate identification, and the VITEK 2 GN09 was used to test the antimicrobial susceptibilities of all isolates. Carbapenem resistance was verified by Etest or K-B diffusion method. Minimal inhibitory concentrations (MICs) of polymyxin B and tigecycline were determined by broth microdilution method for the 18 isolates. The resistance results of tigecycline were interpreted following the European Committee on Antimicrobial Susceptibility Testing (EUCAST) guidelines. Susceptibilities of the other drugs were determined by the criteria of Clinical and Laboratory Standards Institute (CLSI). All methods were carried out in accordance with relevant guidelines and regulations.
4
Carbapenem-Resistant K. pneumoniae Surveillance
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We collected consecutive non-replicate clinical isolates of carbapenem-resistant K. pneumoniae (CRKP) isolated between January 2016 and December 2017 from inpatients at the Shenzhen People’s Hospital, a medical center with 2500 beds in Luohu, Shenzhen, China. Species identification and antimicrobial susceptibility testing were performed with the VITEK-2 compact system (bioMérieux, Marcy-l’Étoile, France). Results were interpreted in accordance with guidelines published by the Clinical and Laboratory Standards Institute (CLSI; document M100-S26).12 The species identifications of all isolates were confirmed with matrix-assisted laser desorption/ionization mass spectrometry (MALDI-TOF-MS, bioMérieux, Marcy-l’Étoile, France). We used the modified Hodge test, performed according to CLSI 2016 recommendations, to identify the CPKP phenotype.12
5
Colistin-Resistant K. pneumoniae Characterization
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The study was conducted at Sanjay Gandhi Postgraduate Institute of Medical Sciences (Lucknow, India), a 900 bed tertiary care referral hospital in North India. Twenty-two colistin resistant K. pneumoniae isolates recovered from various clinical samples like pus, blood, endotracheal aspirate, tissue, and sputum between October 2016 and March 2017 were included in the study. All the isolates were identified using biochemical tests and MALDI-TOF MS (BioMérieux, Marcy l’Étoile, France). Prior to testing, all the isolates were stored in brain heart infusion broth (Becton, Dickinson and Company, Sparks, USA) supplemented with 20% glycerol (Sigma-Aldrich, MO, USA) at −80°C.
6
Bacterial Isolation and Identification
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Bacterial isolation followed the standard microbiological methods [21 ]. We cultured NP/OP swabs and tracheal aspirate samples on sheep blood agar (Oxoid, United Kingdom) and MacConkey (Oxoid, United Kingdom), with an overnight incubation at 37 °C. Blood culture bottles were incubated in BACT/ALERT® VIRTUO 3D Microbial Detection Systems (bioMérieux, Marcy l'Etoile, France), followed by sub-culture for the positive samples onto chocolate blood agar (CBA) (Oxoid, United Kingdom), sheep blood agar (Oxoid, United Kingdom) and MacConkey (Oxoid, United Kingdom). After subculture, we incubated the plates in ambient air; and 5–10% CO2 at 37 °C overnight, followed by isolates' identification using VITEK Matrix-assisted laser desorption ionization–time-of-flight mass spectrometry (MALDI-TOF MS) (bioMérieux, Marcy l'Etoile, France). For quality control, we used Escherichia coli ATCC 8739. All the GNB isolates were transported to the Centre for Microbiology Research, Kenya Medical Research Institute (CMR-KEMRI) laboratories for further analysis.
7
Colistin and Chrysin Efficacy Against A. baumannii
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A total of five non-duplicated COL-R and two colistin-sensitive (COL-S) A. baumannii isolates were recovered from the First Affiliated Hospital of Wenzhou Medical University in China. All these strains were identified by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF/MS; bioMérieux, Lyons, France). ATCC 19606 served as a quality control strain in this study. These strains were conserved in Luria Bertani (LB) broth medium containing 30% glycerol at −80°C until use. All bacterial strains were routinely grown on Luria–Bertani agar plates or LB broth at 37°C before conducting these experiments. Colistin was purchased from Wenzhou Kangtai Biological Technology Co., Ltd. (Zhejiang, China), and chrysin was purchased from MedChemExpress (MCE) Co., Ltd. (NJ, USA). Colistin and chrysin were dissolved in double-distilled water (ddH2O) and diluted with the culture medium to the desired concentrations for in vitro studies. Colistin and chrysin were dissolved in ddH2O and diluted with normal saline to the required concentration for in vivo studies.
8
Yeast Isolation and Antifungal Susceptibility
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For the isolation of yeasts, specimens were inoculated on Sabouraud dextrose agar supplemented with chloramphenicol (50 mg/mL) (Oxoid Limited, Hampshire, UK) and incubated at 36 °C for 72 h. The isolates were identified at the species level by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) (BioMérieux, Marcy L’ Etoile, France; version 3.2). Susceptibility to antifungal agents was determined using the Vitek 2 system (BioMérieux).
9
Isolating and Identifying E. coli from UTI
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A total of 200 non-repetitive E. coli strains isolated from the urine samples of urinary tract infection (UTI) patients admitted to the Affiliated Hospital of Wenzhou Medical University in Wenzhou, China in 2018 were collected. The isolates were identified by using the Matrix-Assisted Laser Desorption/Ionization Time of Flight Mass Spectrometry (MALDI-TOF MS; bioMérieux, Lyons, France).
10
Isolation and Identification of Enterococcus Species
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Swab samples were homogenized in a blender (Waring, New Hartford, Conn.) with 90 ml of buffered peptone water (BPW) (Lab M, Lancashire, UK). After incubation at 37 °C for 24 h, 0.1 ml was streaked onto Slanetz and Bartley Agar (Lab M, Lancashire, UK) and incubated for 24 ± 2 h at 37 ± 1 °C under the same condition 3 . After incubation period, pink or dark red colonies with a narrow, whitish border were observed. After the incubation period, five colonies that showed characteristics of Enterococcus species were selected from each petri dish and transferred to Tryptone Soya Agar (Lab M, Lancashire, UK) agar for purification. The agar plates were then incubated at a temperature of 37 ± 1 °C for 24 ± 2 h. The suspected isolates were biochemically identified using Gram staining and catalase activity. All strains were stored in skimmed milk powder stocks at -80 °C for further testing 22 . The Enterococcus species were identified through MALDI-TOF MS (BioMérieux Inc., Marcy l'Etoile, France), which was performed only on Gram-positive and catalase-negative cocci 23 .
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