Identification and antimicrobial susceptibility testing of microorganisms from positive blood cultures by a combined lysis-centrifugation method with MALDI-TOF MS and VITEK2 System

Authors

  • Donatella Maria Rodio Dep. Public Health and Infectious Diseases, Sapienza University, Rome
  • Filomena Febbraro Department of Pediatrics, “Sapienza” University Rome
  • Gianluca Puggioni Department of Clinical Medicine, “Sapienza” University Rome
  • Camilla Paradisi Department of Public Health and Infectious Diseases, “Sapienza” University Rome
  • Flavia Stangherlin Department of Public Health and Infectious Diseases, “Sapienza” University Rome
  • Carla Prezioso Department of Public Health and Infectious Diseases, “Sapienza” University Rome
  • Guido Antonelli Department of Molecular Medicine and Pasteur Institute-Cenci Bolognetti Foundation, “Sapienza” University Rome
  • Maria Trancassini Department of Public Health and Infectious Diseases, “Sapienza” University Rome
  • Valeria Pietropaolo Dep. Public Health and Infectious Diseases, Sapienza University, Rome

Keywords:

Bacteremia, LCM, MALDI-TOF MS, VITEK®2, AST

Abstract

Background: Rapid identification and the application of antimicrobial susceptibility testing (AST) to microorganisms causing bloodstream infections is pivotal to guide antimicrobial therapy. This study aims to: 1) utilize the Lysis-Centrifugation Method (LCM) not only for identification of microorganisms from positive blood culture bottles, but also for direct AST full panel by Vitek®2 system (bioMérieux, Inc. France) and by disc diffusion plate (Kirby Bauer Method) and 2) analyze the accuracy of these combined methods.

Methods: 124 mono-microbial positive blood culture bottles were included in this study. An aliquot was subjected to LCM and used for the identification by the MALDI-TOF System. Moreover the microbial pellet was used for direct AST testing full panel by VITEK®2  system and by Kirby Bauer Method.

Results: 123 isolates were correctly identified to the species level and 1 isolate was identified to the genus level. Comparing the two utilized AST methods, it was observed that Gram-positive isolates showed an agreement rate of 96.6% (58/60). Enterococcus faecalis was the only microorganism with a major error rate of 0.6% (2/324) related to erythromycin. Among the Gram-negative, the overall agreement rate was 93.3 (56/60). Klebsiella pneumoniae, Escherichia coli and Enterobacter spp. were the major cause of minor error rates (0.6%, 4/709) and major error rates (1.1%, 8/709). Among the yeasts, results showed an agreement rate of 100% (4/4).

Conclusions: Our simple and cost-effective sample preparation method is very useful for rapid identification as well as AST of microorganisms directly from positive blood culture bottles in a clinical setting.

 

DOI: 10.21276/APALM.1212

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Published

12-04-2017

How to Cite

1.
Rodio DM, Febbraro F, Puggioni G, Paradisi C, Stangherlin F, Prezioso C, et al. Identification and antimicrobial susceptibility testing of microorganisms from positive blood cultures by a combined lysis-centrifugation method with MALDI-TOF MS and VITEK2 System. Ann of Pathol and Lab Med [Internet]. 2017 Apr. 12 [cited 2025 Feb. 5];4(2):A170-177. Available from: https://pacificejournals.com/journal/index.php/apalm/article/view/apalm1212

Issue

Vol. 4 No. 2 (2017)

Section

Original Article

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Copyright (c) 2017 Donatella Maria Rodio, Filomena Febbraro, Gianluca Puggioni, Camilla Paradisi, Flavia Stangherlin, Carla Prezioso, Guido Antonelli, Maria Trancassini, Valeria Pietropaolo

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