Abstract

BackgroundRapid identification and antimicrobial susceptibility testing (AST) of the causative agent(s) of bloodstream infections can lead to prompt appropriate antimicrobial therapy. To shorten species identification, in this study bacteria were recovered from monomicrobial blood cultures by serum separator tubes and spotted onto the target plate for direct MALDI-TOF MS identification. Proper antibiotics were selected for direct AST based on species identification. In order to obtain rapid AST results, bacteria were recovered from positive blood cultures by two different protocols: by serum separator tubes (further referred to as PR1), or after a short-term subculture in liquid medium (further referred to as PR2). The results were compared with those obtained by the method currently used in our laboratory consisting in identification by MALDI-TOF and AST by Vitek 2 or Sensititre on isolated colonies.ResultsThe direct MALDI-TOF method concordantly identified with the current method 97.5 % of the Gram-negative bacteria and 96.1 % of the Gram-positive cocci contained in monomicrobial blood cultures. The direct AST by PR1 and PR2 for all isolate/antimicrobial agent combinations was concordant/correct with the current method for 87.8 and 90.5 % of Gram-negative bacteria and for 93.1 and 93.8 % of Gram-positive cocci, respectively. In particular, 100 % categorical agreement was found with levofloxacin for Enterobacteriaceae by both PR1 and PR2, and 99.0 and 100 % categorical agreement was observed with linezolid for Gram-positive cocci by PR1 and PR2, respectively. There was no significant difference in accuracy between PR1 and PR2 for Gram-negative bacteria and Gram-positive cocci.ConclusionsThis newly described method seems promising for providing accurate AST results. Most importantly, these results would be available in a few hours from blood culture positivity, which would help clinicians to promptly confirm or streamline an effective antibiotic therapy in patients with bloodstream infections.Electronic supplementary materialThe online version of this article (doi:10.1186/s12866-016-0805-5) contains supplementary material, which is available to authorized users.

Highlights

  • Rapid identification and antimicrobial susceptibility testing (AST) of the causative agent(s) of bloodstream infections can lead to prompt appropriate antimicrobial therapy

  • In order to adapt the system to rapid AST from positive blood cultures, two different protocols, protocol 1 (PR1) and protocol 2 (PR2), were evaluated, the first using bacteria harvested from positive BC by serum separator tubes, the second using a short-term subculture of a positive BC in liquid medium

  • Identification of bacteria from positive BC by direct MALDI-TOF A total of 194 positive BC were apparently monomicrobial at the Gram staining, eleven (5.7 %) of which resulted to be polymicrobial after subculture and were excluded from this study

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Summary

Introduction

Rapid identification and antimicrobial susceptibility testing (AST) of the causative agent(s) of bloodstream infections can lead to prompt appropriate antimicrobial therapy. In this study bacteria were recovered from monomicrobial blood cultures by serum separator tubes and spotted onto the target plate for direct MALDI-TOF MS identification. In order to obtain rapid AST results, bacteria were recovered from positive blood cultures by two different protocols: by serum separator tubes (further referred to as PR1), or after a short-term subculture in liquid medium (further referred to as PR2). In order to adapt the system to rapid AST from positive blood cultures, two different protocols, protocol 1 (PR1) and protocol 2 (PR2), were evaluated, the first using bacteria harvested from positive BC by serum separator tubes, the second using a short-term subculture of a positive BC in liquid medium.

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