MALDI-TOF MS based procedure to detect KPC-2 directly from positive blood culture bottles and colonies

Autor: Marcela Alejandra Radice, Roque Figueroa-Espinosa, Gabriel Osvaldo Gutkind, Rubén Barrios, Daniela Cejas, Agustina Costa, Carlos Vay, José Alejandro Di Conza
Rok vydání: 2019
Předmět:
Microbiology (medical)
ANTIMICROBIAL RESISTANCE DETECTION
Klebsiella pneumoniae
Sinapinic acid
medicine.disease_cause
Microbiology
beta-Lactamases
KPC-2
purl.org/becyt/ford/1 [https]
03 medical and health sciences
chemistry.chemical_compound
Bacterial Proteins
Enterobacterales
Escherichia coli
polycyclic compounds
medicine
Humans
MALDI-TOF MS
Blood culture
purl.org/becyt/ford/1.6 [https]
Molecular Biology
Escherichia coli Infections
030304 developmental biology
0303 health sciences
biology
medicine.diagnostic_test
030306 microbiology
BLOOD CULTURE
bacterial infections and mycoses
biology.organism_classification
Bacterial Typing Techniques
Klebsiella Infections
Matrix-assisted laser desorption/ionization
chemistry
Blood culture bottles
Spectrometry
Mass
Matrix-Assisted Laser Desorption-Ionization
Positive blood culture
Zdroj: CONICET Digital (CONICET)
Consejo Nacional de Investigaciones Científicas y Técnicas
instacron:CONICET
ISSN: 0167-7012
Popis: In this study, we identified specific carbapenemase-producing isolates applying an easy and rapid protocol for the detection of mature KPC-2 β-lactamase by MALDI-TOF MS from colony and positive blood culture bottles. In addition, we evaluated the correlation of the ~11,109 Da signal as a biomarker associated with KPC-2 production. A collection of 126 well-characterized clinical isolates were evaluated (including 60 KPC-2-producing strains). Presence of KPC-2 was assessed by MALDI-TOF MS on protein extracts. Samples were prepared using the double layer sinapinic acid technique. In order to identify mature KPC-2, raw spectra were analyzed focusing on the range between m/z 25,000–30,000 Da. A single distinctive peak, at approximately m/z 28,544 Da was found in all clinical and control KPC-2-producing strains, and consistently absent in the control groups (ESBL producers and susceptible strains). This peak was detected in all species independently of where the gene bla KPC-2 was embedded. Statistical results showed 100% sensitivity, CI95%: [94.0%; 100%] and 100% specificity, CI95%: [94.6%; 100%], indicating a promising test with a high discriminative power. KPC-2 β-lactamase could be directly detected from both colonies and blood culture bottles. On the other hand, the m/z 11,109 Da signal determinant was only associated with 32% of Klebsiella pneumoniae and Escherichia coli KPC positive isolates. This MALDI-TOF MS methodology has the potential to detect directly the widespread and clinically relevant carbapenemase, KPC-2, in Enterobacterales with a straightforward, low cost process, assuming MALDI-TOF MS is already adopted as the main identification tool, with clear clinical implications on antibiotic stewardship for early infection treatment. Fil: Figueroa Espinosa, Roque Arnulfo. Universidad de Buenos Aires. Facultad de Farmacia y Bioquímica; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Houssay; Argentina Fil: Costa, Agustina. Universidad de Buenos Aires. Facultad de Farmacia y Bioquímica; Argentina Fil: Cejas, Daniela. Universidad de Buenos Aires. Facultad de Farmacia y Bioquímica; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Houssay; Argentina Fil: Barrios, Rubén. No especifíca; Fil: Vay, Carlos Alberto. Universidad de Buenos Aires. Facultad de Farmacia y Bioquímica. Departamento de Bioquímica Clínica; Argentina. Universidad de Buenos Aires. Facultad de Medicina. Hospital de Clínicas General San Martín; Argentina Fil: Radice, Marcela Alejandra. Universidad de Buenos Aires. Facultad de Farmacia y Bioquímica; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Houssay; Argentina Fil: Gutkind, Gabriel Osvaldo. Universidad de Buenos Aires. Facultad de Farmacia y Bioquímica; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Houssay; Argentina Fil: Di Conza, José Alejandro. Universidad de Buenos Aires. Facultad de Farmacia y Bioquímica; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Houssay; Argentina
Databáze: OpenAIRE
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