Detection of KPC directly from positive blood cultures by MALDI-TOF: From research to the clinical microbiology laboratory routine.

Moreira, NK ; Wilhelm, CM ; et al.

In: Journal of microbiological methods, Jg. 221 (2024-06-01), S. 106940

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Bloodstream infections (BSI) caused by carbapenem-resistant Gram-negative bacilli (CR-GNB) are a subject of major clinical concern, mainly those associated with carbapenemase-producing isolates. Matrix-assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-TOF MS) has been proposed to detect specific β-lactamases, including KPC. We aimed to detect KPC enzyme directly from positive blood cultures using MALDI-TOF MS. Overall, 146 clinical Gram-negative bacilli (46 CR-GNB) recovered from consecutive blood cultures were evaluated. Proteins were extracted using formic acid, isopropyl alcohol, and water and spotted onto a steel target plate using the double-layer sinapinic acid method. The relative ions intensity ≥120 arbitrary units (a.u.) of a peak close to 28,700 m/z indicated the presence of KPC. The results were compared to HRM-qPCR methodology. This specific peak was observed in 11/14 blood bottles with bla KPC positive isolates (78.6% sensitivity), with 3 false-positive results (97.7% specificity). Analysis from colonies reached identical sensitivity (78.6%), but higher specificity (100%). The detection of KPC peaks directly from positive blood cultures using MALDI-TOF MS is feasible and rapid. It's excellent specificity indicates that positive results are consistently associated with the presence of a KPC producer in positive blood culture.

Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper.

Titel:	Detection of KPC directly from positive blood cultures by MALDI-TOF: From research to the clinical microbiology laboratory routine.
Autor/in / Beteiligte Person:	Moreira, NK ; Wilhelm, CM ; Collar, GDS ; Echevarria, AD ; Becker, J ; Barth, AL ; Caierão, J
Link:	Zum Volltext
Zeitschrift:	Journal of microbiological methods, Jg. 221 (2024-06-01), S. 106940
Veröffentlichung:	Amsterdam, The Netherlands : Elsevier Biomedical, c1983-, 2024
Medientyp:	academicJournal
ISSN:	1872-8359 (electronic)
DOI:	10.1016/j.mimet.2024.106940
Schlagwort:	Humans Sensitivity and Specificity Gram-Negative Bacteria isolation & purification Gram-Negative Bacteria enzymology Gram-Negative Bacteria classification Gram-Negative Bacteria genetics Bacteremia microbiology Bacteremia diagnosis Gram-Negative Bacterial Infections microbiology Gram-Negative Bacterial Infections diagnosis Gram-Negative Bacterial Infections blood Anti-Bacterial Agents pharmacology Carbapenems pharmacology Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization methods beta-Lactamases genetics Blood Culture methods Bacterial Proteins genetics
Sonstiges:	Nachgewiesen in: MEDLINE Sprachen: English Publication Type: Journal Article Language: English [J Microbiol Methods] 2024 Jun; Vol. 221, pp. 106940. <i>Date of Electronic Publication: </i>2024 May 01. MeSH Terms: Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization* / methods ; beta-Lactamases* / genetics ; Blood Culture* / methods ; Bacterial Proteins* / genetics ; Humans ; Sensitivity and Specificity ; Gram-Negative Bacteria / isolation & purification ; Gram-Negative Bacteria / enzymology ; Gram-Negative Bacteria / classification ; Gram-Negative Bacteria / genetics ; Bacteremia / microbiology ; Bacteremia / diagnosis ; Gram-Negative Bacterial Infections / microbiology ; Gram-Negative Bacterial Infections / diagnosis ; Gram-Negative Bacterial Infections / blood ; Anti-Bacterial Agents / pharmacology ; Carbapenems / pharmacology Contributed Indexing: Keywords: Blood culture; KPC; MALDI-TOF MS Substance Nomenclature: 0 (carbapenemase) Entry Date(s): Date Created: 20240503 Date Completed: 20240517 Latest Revision: 20240517 Update Code: 20240518

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