Detection of carbapenemase activity directly from blood culture vials using MALDI-TOF MS: a quick answer for the right decision

Carvalhaes, Cecília G; Cayô, Rodrigo; Visconde, Marina Francisco; Barone, Talita; Frigatto, Eliete A. M.; Okamoto, Débora N.; Assis, Diego M.; Juliano, Luiz; Machado, Antonia M O; Gales, Ana Cristina

doi:10.1093/jac/dku094

Cited by 66 publications

(46 citation statements)

References 22 publications

(16 reference statements)

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“…Overnight growth of colonies would still be required to identify organisms not identifiable by this method (e.g., polymicrobial specimens, Gram-positive rods, anaerobes, and yeast) and to perform susceptibility testing. That being said, methods have been developed using pelleted bacteria from positive blood cultures to assess antimicrobial resistance by MALDI-TOF MS (22,23). A valuable future study would be to employ our pellet method in parallel with a method for the rapid detection of antimicrobial resistance.…”

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confidence: 99%

Rapid Identification of Positive Blood Cultures by Matrix-Assisted Laser Desorption Ionization–Time of Flight Mass Spectrometry Using Prewarmed Agar Plates

et al. 2014

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bThis study describes an inexpensive and straightforward method for identifying bacteria by matrix-assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF MS) directly from positive blood cultures using prewarmed agar plates. Different inoculation methods and incubation times were evaluated to determine the optimal conditions. The two methods using pelleted material from positive culture bottles performed best. In particular, the pellet streak method correctly identified 94% of the Gram negatives following 4 h of incubation and 98% of the Gram positives following 6 h of incubation.

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confidence: 99%

Rapid Identification of Positive Blood Cultures by Matrix-Assisted Laser Desorption Ionization–Time of Flight Mass Spectrometry Using Prewarmed Agar Plates

et al. 2014

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“…A specific antimicrobial resistance gene can be detected by real-time polymerase chain reaction (RT-PCR), microarray assay or fluorescence in situ hybridization (FISH) and beta-lactamases or carbapenemases have been detected using MALDI-TOF [11,12]. In MDR species such as MRSA, VRE, MDR Enterobacteriaceae, MDR Pseudomonas and Acinetobacter, WGS detects an antimicrobial resistome such as ESBLs or carbapenemases by SNP-level analysis, contributing to antimicrobial therapy.…”

Section: Study Of Antimicrobial Resistancementioning

confidence: 99%

“…When observing another method for the detection of carbapenemase, isolates were incubated with ertapenem; the results were considered positive revealing carbapenem hydrolysis when the peak at m/z 475 disappeared completely [12,50].…”

Section: Species Identification In 3 H Culturementioning

confidence: 99%

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Recent Technology of Clinical Microbiology; Whole Genome Sequencing (WGS) and Matrix-Assisted, Laser Desorption Ionization-Time of Flight Mass Spectrometry (MALDI-TOF MS)

Jeongsook¹

2017

J Microb Biochem Technol

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“…The detection of different classes of carbapenemase activity has been assessed by matrix-assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF MS) (1)(2)(3)(4). In this study, we evaluated the influence of distinct growth culture media on detection of carbapenemase activity by MALDI-TOF MS using Vitek MS equipment (bioMérieux, Brazil).…”

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Influence of Culture Media on Detection of Carbapenem Hydrolysis by Matrix-Assisted Laser Desorption Ionization–Time of Flight Mass Spectrometry

et al. 2016

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In this study, we evaluated the influence of distinct bacterial growth media on detection of carbapenemase hydrolysis by matrixassisted laser desorption ionization-time of flight mass spectrometry. False-negative results were observed for OXA-25-, OXA-26-, and OXA-72-producing Acinetobacter baumannii isolates grown on MacConkey agar medium. The other culture media showed 100% sensitivity and 100% specificity for detecting carbapenemase. Rapid detection of carbapenemase-producing isolates is important for clinical management of infected patients and implementation of infection control measures. The detection of different classes of carbapenemase activity has been assessed by matrix-assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF MS) (1-4). In this study, we evaluated the influence of distinct growth culture media on detection of carbapenemase activity by MALDI-TOF MS using Vitek MS equipment (bioMérieux, Brazil).(This study was presented in part as poster D-1532 at the 54th Interscience Conference on Antimicrobial Agents and Chemotherapy, Washington, DC, 5 to 9 September 2014 [5].)A total of 61 carbapenemase-producing and 32 non-carbapenemase-producing Gram-negative clinical isolates previously characterized by PCR and DNA sequencing were grown on MuellerHinton agar (MHA), blood agar (BA), MacConkey agar (MAC), and chromID CPS agar (CPS) (bioMérieux, Brazil). Sixty-one carbapenemase-producing clinical isolates resistant to at least one carbapenem compound were evaluated, including Klebsiella pneumoniae producers of IMP-1 (n ϭ 6), KPC-2 (n ϭ 25), NDM-1 (n ϭ 1), and BKC-1 (n ϭ 2), Enterobacter cloacae producers of VIM-1 (n ϭ 1), IMP-1 (n ϭ 1), and KPC-2 (n ϭ 1), KPC-2-producing Escherichia coli (n ϭ 1), Serratia marcescens producers of KPC-2 (n ϭ 13) and IMP-1 (n ϭ 1), SPM-1-producing Pseudomonas aeruginosa (n ϭ 5), and Acinetobacter baumannii producers of OXA-25 (n ϭ 1), OXA-26 (n ϭ 1), OXA-72 (n ϭ 1), and OXA-58 (n ϭ 1). In addition, 32 non-carbapenemase-producing bacterial isolates, including Enterobacteriaceae (n ϭ 29), P. aeruginosa (n ϭ 1), and A. baumannii (n ϭ 2) isolates, were tested as negative controls. The MALDI-TOF MS assay was performed by incubating a 1-l loop of fresh bacterial colonies from MHA, BA, MAC, or CPS in 100 l of buffer-adjusted solution (20 mM Tris-HCl [pH 6.8]) with 0.25 mg/ml ertapenem (ETP) (Merck Sharp & Dohme, NJ) (6). The samples were analyzed after 2 and 4 h of incubation at 37°C. One microliter of the supernatant was spotted onto a MALDI target plate, followed by 1 l of ␣-cyano-4-hydroxycinnamic acid (HCCA) matrix solution (bioMérieux) on each spot, and allowed to dry at room temperature. The mass spectrum was obtained by using the Vitek MS instrument operating in linear positive-ion mode and using SARAMIS version 4.1.2 software (bioMérieux). For each isolate, mass spectra were acquired by accumulating 200 laser shots at 50% to 55% laser power in the m/z range of 400 to 600 Da after instrument calibration using the HCCA matrix peaks ( ϩ ) at 497 m...

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Detection of carbapenemase activity directly from blood culture vials using MALDI-TOF MS: a quick answer for the right decision

Abstract: The MALDI-TOF MS carbapenemase assay is a feasible and rapid test to identify carbapenemase activity directly from blood culture vials. It may contribute to faster readjustment of empirical antimicrobial therapy and implementation of infection control measures.

Cited by 66 publications

References 22 publications

Rapid Identification of Positive Blood Cultures by Matrix-Assisted Laser Desorption Ionization–Time of Flight Mass Spectrometry Using Prewarmed Agar Plates

Rapid Identification of Positive Blood Cultures by Matrix-Assisted Laser Desorption Ionization–Time of Flight Mass Spectrometry Using Prewarmed Agar Plates

Recent Technology of Clinical Microbiology; Whole Genome Sequencing (WGS) and Matrix-Assisted, Laser Desorption Ionization-Time of Flight Mass Spectrometry (MALDI-TOF MS)

Influence of Culture Media on Detection of Carbapenem Hydrolysis by Matrix-Assisted Laser Desorption Ionization–Time of Flight Mass Spectrometry

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