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Ultra-rapid flow cytometry assay for colistin MIC determination in Enterobacterales, Pseudomonas aeruginosa and Acinetobacter baumannii

2020Journal article Open access
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dc.contributor.authorFonseca e Silva, Daniela
dc.contributor.authorAndrade, Ferdinando F.
dc.contributor.authorGomes, Rosário
dc.contributor.authorSilva-Dias, Ana
dc.contributor.authorMartins-Oliveira, Inês
dc.contributor.authorPérez-Viso, Blanca
dc.contributor.authorRamos, Maria Helena
dc.contributor.authorRodrigues, Acácio G.
dc.contributor.authorCantón, Rafael
dc.contributor.authorPina-Vaz, Cidália
dc.date.accessioned2021-11-16T13:31:04Z
dc.date.available2021-11-16T13:31:04Z
dc.date.issued2020
dc.description.abstractObjectives: Both EUCAST and CLSI recommend broth microdilution for antimicrobial susceptibility testing of colistin, but this method is cumbersome and takes 16-24 h to give results. Our objective was to evaluate a rapid quantitative colistin MIC susceptibility assay based on flow cytometry analysis (FASTcolistin MIC) in comparison with standard broth microdilution assay. Methods: One hundred and sixteen Gram-negative bacilli (78 Enterobacterales, 28 Pseudomonas aeruginosa and 10 Acinetobacter baumannii) were studied in parallel using standard broth microdilution following EUCAST recommendations and FASTcolistin MIC kit. In the last one, a bacteria suspension (0.5 MacFarland) was prepared, diluted in Muller-Hinton broth, incubated in the susceptibility panel containing different colistin concentrations (range 0.125-64 mg/L) with a fluorescent probe and incubated 1 h at 35ºC. After that, a flow cytometry analysis using CytoFLEX (Beckmam) was performed. Using a dedicated software (BioFAST) an automated MIC result was obtained after 1.5 h. Performance evaluation was performed according to the ISO standard 20776-2. Reproducibility and repeatability, categorical (CA) and essential agreement (EA), and lot-to-lot variation and operator-to-operator variability, as well as time to results were determined. Results: Overall, 100% CA (CI 97-100%) and 95.7% EA (CI 90-98%) was obtained with high repeatability (100%; CI 80-100%)and reproducibility (97%; (CI 83-99%)). Absence of lot-to-lot variations or differences in the operators' performance was observed. Conclusions: FASTcolistin MIC is an accurate, reliable and ultra-rapid method (1 h incubation versus 24 h) for susceptibility testing of colistin of common Gram-negative bacilli recovered in clinical laboratories.pt_PT
dc.description.versioninfo:eu-repo/semantics/publishedVersionpt_PT
dc.identifier.citationFonseca E Silva D, Andrade FF, Gomes R, et al. Ultra-rapid flow cytometry assay for colistin MIC determination in Enterobacterales, Pseudomonas aeruginosa and Acinetobacter baumannii. Clin Microbiol Infect. 2020;26(11):1559.e1-1559.e4. doi:10.1016/j.cmi.2020.08.019pt_PT
dc.identifier.doi10.1016/j.cmi.2020.08.019pt_PT
dc.identifier.issn1469-0691
dc.identifier.issn1198-743X
dc.identifier.urihttp://hdl.handle.net/10400.16/2557
dc.language.isoengpt_PT
dc.peerreviewedyespt_PT
dc.publisherElsevierpt_PT
dc.relation.publisherversionhttps://www.clinicalmicrobiologyandinfection.com/article/S1198-743X(20)30502-4/fulltextpt_PT
dc.subjectASTpt_PT
dc.subjectAntibiotic susceptibilitypt_PT
dc.subjectColistinpt_PT
dc.subjectFlow-cytometrypt_PT
dc.subjectMDR microorganismspt_PT
dc.subjectMIC determinationpt_PT
dc.subjectMicrodilutionpt_PT
dc.titleUltra-rapid flow cytometry assay for colistin MIC determination in Enterobacterales, Pseudomonas aeruginosa and Acinetobacter baumanniipt_PT
dc.typejournal article
dspace.entity.typePublication
oaire.citation.conferencePlaceEnglandpt_PT
oaire.citation.endPage1559.e4pt_PT
oaire.citation.issue11pt_PT
oaire.citation.startPage1559.e1pt_PT
oaire.citation.titleClinical Microbiology and Infectionpt_PT
oaire.citation.volume26pt_PT
rcaap.rightsopenAccesspt_PT
rcaap.typearticlept_PT

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