Evaluation of antimicrobial susceptibility testing of Nocardia spp. isolates by broth microdilution with resazurin and spectrophotometry.

Vivian Caso Coelho, Samara D'Ajuda Pereira Neves, Mauro Cintra Giudice, Gil Benard, Marta Heloisa Lopes, Paula Keiko Sato
Author Information
  1. Vivian Caso Coelho: Laboratory of Medical Investigation in Immunology (LIM48), Hospital das Clinicas HCFMUSP, Faculdade de Medicina, Universidade de Sao Paulo, Av. Dr. Eneas Carvalho de Aguiar 470, IMT 2, terreo, Sao Paulo, SP, 05403-000, Brazil.
  2. Samara D'Ajuda Pereira Neves: Programa de Aprimoramento Profissional, HCFMUSP (LIM48), Sao Paulo, Brazil.
  3. Mauro Cintra Giudice: Laboratory of Medical Investigation in Micology (LIM53), HCFMUSP, Sao Paulo, Brazil.
  4. Gil Benard: Laboratory of Medical Investigation in Micology (LIM53), HCFMUSP, Sao Paulo, Brazil.
  5. Marta Heloisa Lopes: Laboratory of Medical Investigation in Immunology (LIM48), Hospital das Clinicas HCFMUSP, Faculdade de Medicina, Universidade de Sao Paulo, Av. Dr. Eneas Carvalho de Aguiar 470, IMT 2, terreo, Sao Paulo, SP, 05403-000, Brazil.
  6. Paula Keiko Sato: Laboratory of Medical Investigation in Immunology (LIM48), Hospital das Clinicas HCFMUSP, Faculdade de Medicina, Universidade de Sao Paulo, Av. Dr. Eneas Carvalho de Aguiar 470, IMT 2, terreo, Sao Paulo, SP, 05403-000, Brazil. paula.s@hc.fm.usp.br. ORCID

Abstract

BACKGROUND: Nocardia species are ubiquitous in natural environments and can cause nocardiosis. In the present study, the use of Resazurin salt and Spectrophotometry were proposed as alternative methods to reduce subjectivity in the interpretation of susceptibility results to antimicrobials by the broth microdilution method for Nocardia spp.
RESULTS: The susceptibility of Nocardia spp. isolates to Amikacin, Ciprofloxacin, Minocycline and Trimethoprim-Sulfamethoxazole was evaluated by Minimum Inhibitory Concentration (MIC) determinations by the broth microdilution method. To verify cellular growth, the colour-changing dye Resazurin was applied, the Optical Densities were measured on a spectrophotometer, and both were compared to Clinical and Laboratory Standards Institute (CLSI) Gold Standard method (visual MIC determination). Percentages of essential and categorical agreements and interpretative categorical errors were calculated within each method (intra-reading) and between them (inter-reading). The Gold Standard visual reading demonstrated 100% of essential and categorical intra-reading agreements for Amikacin, and there was no error when compared with the alternative methods. For Ciprofloxacin, the comparison between the Gold Standard and the Spectrophotometric reading showed 91.5% of essential agreement. In the categorical intra-reading analysis for Minocycline, there were 88.1 and 91.7% in the Gold Standard and in the Spectrophotometric readings, respectively, and 86.4% of concordance between them. High rates of categorical agreement were also observed on the Trimethoprim-Sulfamethoxazole analyses, with 93.7% for the Gold Standard, 84.9% for the Resazurin readings, and 80.5% between them.
CONCLUSIONS: The alternative methods with Resazurin and Spectrophotometric readings showed high agreement rates with the Gold Standard.

Keywords

References

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MeSH Term

Anti-Bacterial Agents
Humans
Microbial Sensitivity Tests
Nocardia
Nocardia Infections
Oxazines
Spectrophotometry
Xanthenes

Chemicals

Anti-Bacterial Agents
Oxazines
Xanthenes
resazurin

Word Cloud

Created with Highcharts 10.0.0GoldStandardNocardiaResazurincategoricalsusceptibilitymicrodilutionmethodalternativemethodsbrothsppessentialintra-readingSpectrophotometricagreementreadingsSpectrophotometryisolatesAmikacinCiprofloxacinMinocyclineTrimethoprim-SulfamethoxazoleMinimumMICcomparedvisualagreementsreadingshowed915%7%ratestestingBACKGROUND:speciesubiquitousnaturalenvironmentscancausenocardiosispresentstudyusesaltproposedreducesubjectivityinterpretationresultsantimicrobialsRESULTS:evaluatedInhibitoryConcentrationdeterminationsverifycellulargrowthcolour-changingdyeappliedOpticalDensitiesmeasuredspectrophotometerClinicalLaboratoryStandardsInstituteCLSIdeterminationPercentagesinterpretativeerrorscalculatedwithininter-readingdemonstrated100%errorcomparisonanalysis881respectively864%concordanceHighalsoobservedanalyses93849%80themCONCLUSIONS:highEvaluationantimicrobialresazurinspectrophotometryAntimicrobialBrothinhibitoryconcentration

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