Abstract

Matrix-assisted laser desorption ionization time of flight mass spectrometry (MALDI-TOF MS) is a novel method for the direct identification of bacteria from blood culture broths. We evaluate for the first time, the performance of the MALDI Sepsityper™ Kit and MS for the identification of bacteria compared to standard phenotypic methods using the manufacturer's specified bacterial identification criteria (spectral scores ≥1.700–1.999 and ≥2.000 indicated identification to genus and species level, respectively). Five hundred and seven positive blood culture broths were prospectively examined, of which 379 (74.8%; 358 monomicrobial, 21 polymicrobial) were identified by MALDI-TOF MS; 195 (100%) and 132 (67.7%) of 195 gram-positive; and 163 (100%) and 149 (91.4%) of 163 gram-negative organisms from monomicrobial blood cultures were correctly identified to genus and species level, respectively. Spectral scores <1.700 (no identification) were obtained in 128/507 (25.2%) positive blood culture broths, including 31.6% and 32.3% of gram-positive and polymicrobial blood cultures, respectively. Significantly more gram-negative organisms were identified compared to gram-positive organisms at species level (p<0.0001). Five blood cultures were misidentified, but at species level only; including four monomicrobial blood cultures with Streptococcus oralis/mitis that were misidentified as Streptococcus pneumoniae. Positive predictive values for the direct identification of both gram-positive and gram-negative bacteria from monomicrobial blood culture broths to genus level were 100%. A diagnostic algorithm for positive blood culture broths that incorporates gram staining and MALDI-TOF MS should identify the majority of pathogens, particularly to genus level.

Highlights

  • Bloodstream infections (BSIs) are a significant cause of morbidity and mortality in hospitals

  • This study focused on bacteria only; yeasts were excluded from the analysis

  • We report the performance of the MALDI SepsityperTM Kit that provides all the reagents required to process blood culture broths according to a standardized protocol prior to MALDI-TOF MS analysis

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Summary

Introduction

Bloodstream infections (BSIs) are a significant cause of morbidity and mortality in hospitals. Rapid nucleic acid amplification methods such as real-time PCR using melting curve analysis, multiplex PCR, fluorescence in situ hybridization (FISH) and peptide nucleic acidFISH (PNA-FISH) have been used to detect pathogens in blood cultures including Staphylococcus aureus, Enterococcus faecalis and Candida albicans [6,7,8]. These assays, only target specific organisms; require technical expertise; and specimens are usually processed in batches.

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