Beckman Coulter MicroScan Manuel D'utilisation page 3

Candida albicans ATCC 66027
Candida kefyr ATCC 66028
Candida tropicalis ATCC 66029
Cryptococcus albidus ATCC 66030
Cryptococcus neoformans ATCC 66031
Candida glabrata ATCC 66032
Cryptococcus uniguttulatus ATCC 66033
PROCEDURE
Preparation of Panels
1. Remove the panels to be used from storage. Panels should not be used if the desiccant is not present or is broken
or if the integrity of the packaging is compromised (unsealed, punctured, or torn).
2. Cut open the pouch and remove the panel. Remove the panel immediately from the foil pouch. Allow panels to equilibrate
to room temperature prior to rehydration. Panels may be stacked with clean Cover Tray on top.
Inoculum Preparation
1. Inoculate the Rapid Yeast Identification Panel with an actively growing organism. Colonies from a primary isolation
Sabouraud Dextrose Agar plate may be used if sufficient growth is present. If necessary, subculture the yeast isolate
onto a Sabouraud Dextrose Agar plate and incubate until sufficient growth occurs. Incubation for 48 hours at 30°C or 48
to 72 hours at 25°C (room temperature) is recommended.
NOTE: Cultures incubated for more than 48 hours at 30°C should be used only if additional incubation is required to
obtain adequate growth.
2. Using a sterile swab or inoculating loop, remove growth from the agar plate and emulsify in 3 mL of Inoculum Water
(autoclaved deionized water in a 13 x 84 mm or 13 x 100 mm tube). Each suspension must be compared and be
equivalent to the MicroScan Yeast Turbidity Standard. The comparison to the test inoculum suspension and the Yeast
Turbidity Standard should be done using an adequate light source and by comparing the tubes to a white card with
contrasting black lines.
NOTE: Take care to insure that no agar medium is removed with the organism.
3. Mix or vortex the suspension until it is homogeneous. Some yeast strains do not readily disperse in water. If vortexing
does not result in a homogeneous suspension, let the suspension sit for 10-15 minutes and revortex. If the suspension
still has clumps, allow the clumps to settle to the bottom of the tube and use the supernatant to inoculate the yeast panel.
The turbidity of the supernatant must match the MicroScan Yeast Turbidity Standard. If it does not, transfer more growth
to the Inoculum Water and repeat the above procedure.
NOTE: DO NOT pipette clumps of cells into the yeast panel.
Panel Inoculation
1. Label the panel with the specimen number.
2. Add 50 μL of yeast suspension to each substrate-containing well plus the control wells BNAC and NPC. Use of a Pasteur
pipette for inoculation is not recommended.
Table 1-1, Substrates
Substrates
Hydroxyproline-β-Naphthylamide
L-Isoleucine-β-Naphthylamide
L-Proline-β-Naphthylamide
L-Tyrosine-β-Naphthylamide
Glycine β-Naphthylamide
Glycylglycine-β-Naphthylamide
Glycyl-L-Arginine-4-Methoxy-β-Naphthylamide
Glycyl-L-Proline-4-Methoxy-β-Naphthylamide
L-Arginyl-L-Arginine-β-Naphthylamide
L-Lysyl-L-Alanine-4-Methoxy-β-Naphthylamide
L-Alanine-4-Methoxy-β-Naphthylamide
L-Seryl-L-Tyrosine-β-Naphthylamide
Urea
3-Indoxyl Phosphate
1. Different formulations of the same Substrates are used.
* American Type Culture Collection, Manassas, VA USA
C29879–AB
*
Abbr. Substrates
HPR L-Histidine-β-Naphthylamide
ILE Sucrose
PRO Sucrose
TYR Trehalose
GLY p-Nitrophenyl-α-D-Glucopyranoside
GGLY p-Nitrophenyl-α-D-Glucopyranoside
GLAR p-Nitrophenyl-β-D-Glucopyranoside
GLPR o-Nitrophenyl-β-D-Galactopyranoside
AARG p-Nitrophenyl-β-D-Fucopyranoside
LYAL
p-Nitrophenyl-α-D-Galactopyranoside
ALA p-Nitrophenyl-N-Acetyl-β-D-Glucosamine
STY p-Nitrophenyl-β-D-Cellobiose
URE p-Nitrophenyl-N-Acetyl-β-D-Galactosaminide
IDX
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Abbr.
HIS
SUC1
1
SUC2
TRE
AGL1
1
AGL2
BGL
BGAL
BDF
AGAL
NAG
CELL
NGAL