cphm-838 aspergillus species by laser light scattering...
TRANSCRIPT
Rapid Voriconazole Susceptibility Testing ofAspergillus Species by Laser Light Scattering
A. Fitzgerald1, W. Memon1, A. Tomaras2, S.X. Zhang1
1. Johns Hopkins Hospital, Baltimore, MD2. BacterioScan, St. Louis, MO
Introduction
In vitro antifungal susceptibility testing for Aspergillus species is currently performed using the CLSI-developed microbroth dilution
method, which takes at least 48 hours to obtain MIC results. There is an unmet need for a more rapid antifungal susceptibility testing
method for Aspergillus. A laser light scattering system (BacterioScan 216Dx) is able to detect microbial growth in broth by producing
signals based on optical density read every 5 minutes. The purpose of this study was to test the BacterioScan 216Dx system for faster
antifungal susceptibility results compared to the current CLSI microbroth dilution method.
Materials
20 Aspergillus species with previously performed Voriconazole susceptibility via
microbroth dilution method were tested on the BacterioScan 216Dx (Figure 1). The
isolates cultured on Potato Flake Agar were <7 days old at time of testing. Purity plates
(Sabouraud Dextrose) were inoculated from the test cartridge growth control well.
• B-MIC determining timepoint results were available in less than 24 hrs, with a mean of 1060 minutes (17.7
hours) and a median of 1013 minutes (16.9 hours)
• All B-MIC results were concordant to the CLSI MIC results within 2 doubling dilution ranges
• Possible need to determine clinical antifungal breakpoints unique to BacterioScan
• BacterioScan was able to detect the MICs for the 4 Aspergillus fumigatus isolates with an ECV ≥ 1 ug/mL at
a time frame of 669 to 1026 min (11 to 17 h)
• Promising for more rapid mold antifungal testing that will improve patient care and turnaround time
Summary
Acknowledgements:
Thank you to BacterioScan for providing ongoing instrument and reagent support for this project.
Thank you to Dr. Nathan Wiederhold of the Fungus Testing Laboratory, University of Texas at San Antonio for specimens and
genomic information.
Methods
Results
CPHM-8386/22/19
Figure 1
Testing for Optimal Growth Conditions
Organisms: Aspergillus fumigatus, A. niger, A.
flavus, and A. terreus
Medium: Sabouraud Dextrose broth vs. RPMI
broth
% Glucose: 0.2% vs. 2% vs. 4%
Inoculum Dilution/Concentration: 1.0 McFarland
concentration [McF], 0.5 McF, 1:50 dilution of 1.0
McF, 1:50 dilution of 0.5 McF
Best Growth Condition Determined
Medium: RPMI broth
% Glucose: 0.2%
Inoculum Dilution/Concentration: 1:50 dilution of
1.0 McF (OD range 0.18 to 0.22)
• 2 cartridges pre-loaded with Voriconazole
(ug/mL) (See Figure 2) were run for each isolate
• Pipetted 2.5 mL of inoculated RPMI in each well
• Cartridges inoculated at 37°C for 1440 min (24
hr)
• Kinetic laser scattering signals based on growth
were captured and data were converted into
figures.
Figure 2
Cartridge 2
Cartridge 1