Beckman Coulter MicroScan B1016-138 Manuel D'utilisation page 4

PROCEDURE
Panel Preparation
1. Remove the panels to be used from storage. Do not use if the integrity of the packaging is compromised (unsealed,
punctured, or torn).
2. Cut open the pouch and remove the panel. If stored in the refrigerator, remove the panel immediately from the foil pouch.
3. Panels should not be used if any of the following conditions exist:
A. Desiccant is not present or is broken.
B. Panel wells are discolored (e.g., PHO, various antimicrobics).
4. Allow panels to equilibrate to room temperature prior to rehydration. Panels may be stacked with a clean Cover Tray on
top. All opened panels should be used within the same day or discarded.
Inoculum Preparation
CLSI recommends periodically checking your inoculum densities by performing colony counts. Refer to CLSI document
M07-A10 for colony count recommendations. The expected results for E. coli ATCC 25922 should closely approximate 5
x 10 5 CFU/mL for the final test concentrations.
with manual methods that are technique dependent such as the Prompt System or inoculum prepared without the aid of a
photometric device.
Note: Log and Stationary phase techniques are not supported with MicroScan products.
1. Turbidity Standard Technique - Primary Inoculum Method
The turbidity standard technique is recommended for direct inoculation of all aerobic gram positive cocci or for detection
of methicillin resistant staphylococci.
A. Using a sterile applicator stick, loop or swab, touch the surface of 4-5 large or 5-10 small morphologically similar,
well-isolated colonies from an 18-24 hour non-inhibitory agar plate.
B. Emulsify in 3 mL of Inoculum Water (autoclaved deionized water).
C. Cap tightly and vortex the suspension for 2-3 seconds. The final turbidity should be equivalent to that of a 0.5
McFarland Turbidity Standard. An equivalent turbidity can be achieved by using a MicroScan Turbidity Meter with
a range of 0.08 ± 0.02.
D. Pipet 0.1 mL (100 μL) of the standardized suspension into 25 mL of Inoculum Water with PLURONIC. Cap tightly.
Invert 8-10 times to mix.
2. Prompt System
The Prompt System may be used for the more rapidly growing gram positive cocci. Refer to the Prompt Inoculation
procedural manual for the proper use of the Prompt System.
Note: Use with caution. Under-inoculation may occur with organisms that do not meet the size requirement and could
cause incorrect susceptibility and identification test results. Additionally, staphylococci colony counts may be elevated
with the Prompt method and may be greater than the CLSI expected range. Elevated colony counts may adversely affect
antibiotic results that are affected by inoculum. Refer to the Limitations section for antimicrobials which are known to be
inoculum dependent.
Panel Rehydration/Inoculation
Rehydration and inoculation is performed using the RENOK system with Inoculators-D (B1013-4). Refer to RENOK Operator's
Manual for use. If an alternate system is used, rehydrate with 115 ± 10 μL of Inoculum Water (PLURONIC). A final well
concentration of 3-7 X 10
should be prepared by streaking the inoculum to an appropriate agar plate and incubating under the appropriate conditions.
If two or more colony types are present on the purity plate, re-isolate the colonies and retest.
Biochemical Overlays
1. Using a dropper bottle, overlay the ARG and URE wells with at least 3 drops of mineral oil. (These wells are underlined
on the panel.)
2. The media in the wells must be completely covered with mineral oil, but the oil should not overflow the wells.
NOTE: WalkAway SI and WalkAway plus instruments (and WalkAway instruments upgraded with the automated oil
overlay feature) automatically add oil to the appropriate wells.
Incubation
1. Panels can be incubated in a WalkAway System or incubated off-line using the following steps:
A. To ensure even thermal distribution during incubation, stack the panels in groups of 3-5.
C29870–AD
1
CFU/mL should be achieved. To ensure viability and purity of the organism tested, a purity plate
5
The user should pay careful attention to inoculum preparation, especially
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