Rapid identification and antimicrobial susceptibility testing of positive blood cultures using MALDI-TOF MS and a modification of the standardised disc diffusion test: a pilot study.
OBJECTIVE
In an era when clinical microbiology laboratories are under increasing financial pressure, there is a need for inexpensive, yet effective, rapid microbiology tests. The aim of this study was to evaluate a novel modification of standard methodology for the identification and antimicrobial susceptibility testing (AST) of pathogens in positive blood cultures, reducing the turnaround time of laboratory results by 24 h.


METHODS
277 positive blood cultures had a Gram stain performed and were subcultured and incubated at 37°C in a CO2atmosphere for 4-6 h. Identification of the visible growth was performed using matrix-assisted laser desorption time of flight mass spectrometry (MALDI-TOF MS). Taking a modified approach to the Clinical and Laboratory Standards Institute-standardised AST methodology, an inoculum density of 0.5 McFarland was prepared from the early growth for disc diffusion testing. The standard AST method was also performed on the 18-24 h culture.


RESULTS
96% (n=73/76) of gram-negative organisms were correctly identified by MALDI-TOF MS. Comparative analysis of the rapid and standard AST results showed an overall interpretive category error rate of 7.7% (6.7% minor errors, 0.6% major errors and 0.4% very major errors). 100% of Staphylococcus aureus (n=41) and enterococcus isolates (n=9) were correctly identified after 4-6 h incubation. The overall AST categorical agreement was also 100% for these isolates.


CONCLUSIONS
An incubation of 4-6 h directly from positive blood cultures allowed for both a rapid species identification and an antimicrobial susceptibility result approximately 24 h earlier than is possible using standard methodology.
Microbiology Laboratory, University Hospital Waterford, Waterford, Ireland.
C
C
Fitzgerald
Microbiology Laboratory, University Hospital Waterford, Waterford, Ireland.
P
P
Stapleton
Microbiology Laboratory, University Hospital Waterford, Waterford, Ireland.
E
E
Phelan
Microbiology Laboratory, University Hospital Waterford, Waterford, Ireland.
P
P
Mulhare
Microbiology Laboratory, University Hospital Waterford, Waterford, Ireland.
B
B
Carey
Microbiology Laboratory, University Hospital Waterford, Waterford, Ireland.
M
M
Hickey
Microbiology Laboratory, University Hospital Waterford, Waterford, Ireland.
B
B
Lynch
Microbiology Laboratory, University Hospital Waterford, Waterford, Ireland.
M
M
Doyle
ANTIMICROBIAL RESISTANCE; BACTERAEMIA; BLOOD CULTURE; MICROBIOLOGY