Ann Lab Med.  2016 Mar;36(2):117-123. 10.3343/alm.2016.36.2.117.

Direct Identification and Antimicrobial Susceptibility Testing of Bacteria From Positive Blood Culture Bottles by Matrix-Assisted Laser Desorption/Ionization Time-of-Flight Mass Spectrometry and the Vitek 2 System

• Jo SJ
• Park KG
• Han K
• Park DJ
• Park YJ

• KMID: 2373510
• DOI: http://doi.org/10.3343/alm.2016.36.2.117

Abstract

BACKGROUND
We evaluated the reliability and accuracy of the combined use of matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) bacterial identification and Vitek 2 antimicrobial susceptibility testing (AST) for bacteria from positive blood culture bottles.
METHODS
Direct identification and AST were performed in parallel to the standard methods in monomicrobial positive blood culture bottles. In total, 254 isolates grown on aerobic and/or anaerobic bottles were identified with MALDI-TOF Vitek MS (bioMerieux, France), and 1,978 microorganism/antimicrobial agent combinations were assessed. For isolates from anaerobic bottles, an aliquot of the culture broth was centrifuged, washed, and filtered through a nylon mesh. For isolates from aerobic/pediatric bottles, a lysis step using 9.26% ammonium chloride solution and 2% saponin solution was included.
RESULTS
The overall correct identification rate was 81.8% (208/254) and that for gram-positive/gram-negative isolates was 73.9%/92.6%, respectively, and it was 81.8%, 87.6%, and 57.9% for isolates from aerobic, anaerobic, and pediatric bottles, respectively. Identification was not possible in 45 cases, and most of these isolates were streptococci (N=14) and coagulase-negative staphylococci (N=11). Misidentification occurred only in one case. Compared with standard methods, direct AST showed 97.9% (1,936/1,978) agreement with very major error of 0.25%, major error of 0.05%, and minor error of 1.8%.
CONCLUSIONS
This simple and cost-effective sample preparation method gives reliable results for the direct identification and AST of bacteria. For the identification of streptococci and coagulase-negative staphylococci, the method should be further improved.

Keyword

Direct identification; MALDI-TOF MS; Vitek MS; Vitek 2; Positive blood culture

MeSH Terms

Adult
Ammonium Chloride/chemistry
Anti-Infective Agents/*pharmacology
Child
Gram-Negative Bacteria/drug effects/*isolation & purification/metabolism
Gram-Positive Bacteria/drug effects/*isolation & purification/metabolism
Humans
Reagent Kits, Diagnostic
Saponins/chemistry
*Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization
Anti-Infective Agents
Ammonium Chloride
Reagent Kits, Diagnostic
Saponins

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