Rapid detection of bacteria that produce extended-spectrum β-lactamase by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry

ABSTRACT: Objectives: Proper use of antibacterial agents is necessary to prevent the spread of drug-resistant bacteria. To support clinicians, laboratories need to rapidly determine bacterial drug susceptibility/resistance. We have established a method to distinguish extended-spectrum β-lactamase (...

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Autores principales:	Daiki Kaji, Yuriko Matsumura, Atsuo Iwasawa, Satoshi Kimura, Akiko Iwama
Formato:	article
Lenguaje:	EN
Publicado:	Elsevier 2021
Materias:	MALDI-TOF/MS Rapid detection Hydrolysis assay Escherichia coli Klebsiella pneumoniae Proteus mirabilis Microbiology QR1-502
Acceso en línea:	https://doaj.org/article/ec8ad293c0df4914acfaf4d355ee583b
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spelling	oai:doaj.org-article:ec8ad293c0df4914acfaf4d355ee583b2021-11-28T04:32:19ZRapid detection of bacteria that produce extended-spectrum β-lactamase by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry2213-716510.1016/j.jgar.2021.09.009https://doaj.org/article/ec8ad293c0df4914acfaf4d355ee583b2021-12-01T00:00:00Zhttp://www.sciencedirect.com/science/article/pii/S2213716521002319https://doaj.org/toc/2213-7165ABSTRACT: Objectives: Proper use of antibacterial agents is necessary to prevent the spread of drug-resistant bacteria. To support clinicians, laboratories need to rapidly determine bacterial drug susceptibility/resistance. We have established a method to distinguish extended-spectrum β-lactamase (ESBL)-producing clinical isolates by capturing structural changes in β-lactam antibiotics using matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF/MS). Methods: Clinical isolates of Escherichia coli, Klebsiella pneumoniae and Proteus mirabilis, classified into ESBL-producing strains and sensitive strains based on the presence or absence of a CTX-M-type gene, were used. Test bacteria were cultured aerobically in solid-phase wells of Eiken DPD1 dry plates at 35°C for 15 min or 30 min with the antibiotics cefotaxime (CTX), cefpodoxime (CPDX) or piperacillin (PIPC). Culture supernatants were then used for analysis with a MALDI Biotyper. Results: Signals derived from non-hydrolyzed products of antibiotics were observed in all strains. In the case of ESBL-producing strains, signals derived from the hydrolysis products of antibiotics were also observed. Since the ratio of signal intensity derived from hydrolysis products divided by the total signal intensity detected was ≥11% for CTX and ≥6% for CPDX and PIPC, all strains were determined to be ESBL-producing bacteria. Conclusion: The short incubation time of 15 min suggests that this method can identify ESBL-producing strains much more rapidly than conventional methods.Daiki KajiYuriko MatsumuraAtsuo IwasawaSatoshi KimuraAkiko IwamaElsevierarticleMALDI-TOF/MSRapid detectionHydrolysis assayEscherichia coliKlebsiella pneumoniaeProteus mirabilisMicrobiologyQR1-502ENJournal of Global Antimicrobial Resistance, Vol 27, Iss , Pp 309-314 (2021)
institution	DOAJ
collection	DOAJ
language	EN
topic	MALDI-TOF/MS Rapid detection Hydrolysis assay Escherichia coli Klebsiella pneumoniae Proteus mirabilis Microbiology QR1-502
spellingShingle	MALDI-TOF/MS Rapid detection Hydrolysis assay Escherichia coli Klebsiella pneumoniae Proteus mirabilis Microbiology QR1-502 Daiki Kaji Yuriko Matsumura Atsuo Iwasawa Satoshi Kimura Akiko Iwama Rapid detection of bacteria that produce extended-spectrum β-lactamase by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry
description	ABSTRACT: Objectives: Proper use of antibacterial agents is necessary to prevent the spread of drug-resistant bacteria. To support clinicians, laboratories need to rapidly determine bacterial drug susceptibility/resistance. We have established a method to distinguish extended-spectrum β-lactamase (ESBL)-producing clinical isolates by capturing structural changes in β-lactam antibiotics using matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF/MS). Methods: Clinical isolates of Escherichia coli, Klebsiella pneumoniae and Proteus mirabilis, classified into ESBL-producing strains and sensitive strains based on the presence or absence of a CTX-M-type gene, were used. Test bacteria were cultured aerobically in solid-phase wells of Eiken DPD1 dry plates at 35°C for 15 min or 30 min with the antibiotics cefotaxime (CTX), cefpodoxime (CPDX) or piperacillin (PIPC). Culture supernatants were then used for analysis with a MALDI Biotyper. Results: Signals derived from non-hydrolyzed products of antibiotics were observed in all strains. In the case of ESBL-producing strains, signals derived from the hydrolysis products of antibiotics were also observed. Since the ratio of signal intensity derived from hydrolysis products divided by the total signal intensity detected was ≥11% for CTX and ≥6% for CPDX and PIPC, all strains were determined to be ESBL-producing bacteria. Conclusion: The short incubation time of 15 min suggests that this method can identify ESBL-producing strains much more rapidly than conventional methods.
format	article
author	Daiki Kaji Yuriko Matsumura Atsuo Iwasawa Satoshi Kimura Akiko Iwama
author_facet	Daiki Kaji Yuriko Matsumura Atsuo Iwasawa Satoshi Kimura Akiko Iwama
author_sort	Daiki Kaji
title	Rapid detection of bacteria that produce extended-spectrum β-lactamase by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry
title_short	Rapid detection of bacteria that produce extended-spectrum β-lactamase by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry
title_full	Rapid detection of bacteria that produce extended-spectrum β-lactamase by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry
title_fullStr	Rapid detection of bacteria that produce extended-spectrum β-lactamase by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry
title_full_unstemmed	Rapid detection of bacteria that produce extended-spectrum β-lactamase by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry
title_sort	rapid detection of bacteria that produce extended-spectrum β-lactamase by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry
publisher	Elsevier
publishDate	2021
url	https://doaj.org/article/ec8ad293c0df4914acfaf4d355ee583b
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Rapid detection of bacteria that produce extended-spectrum β-lactamase by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry

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