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METHODS OF

EXAMINATION
mycology

Wet Preparation: simplest |


NSS+culture

Temporary Mount
KOH 10-20%: used for specimens
with distracting tissue debris and
cells(vaginal secretions, nails)

Lactophenol Cotton Blue (LPCB):


preserves and stains fungi (shades of
blue) | contains lactic acid (preserve
fungal element), phenol (killing
agent), aniline blue (dye)

India Ink/Nigrossin: negative staining


technique to demonstrate capsule of
C. neoformans

Calcoflour White Stain: uses


fluorescence microscope; Fungal
elements will typically appear bluish
white against dark background

Permanent Mounts
Gram Stain: for detection of yeast
(+)

Giemsa or Wrights: for suspected


cases of Histoplasma capsulatum;
demonstrates yeast cells within
macrophage

Periodic Acid Schiff (PAS): for internal


details of fungal elements

Gomoris methenamine Silver (GMS):


better stain for fungi because it
provides high contrast with minimal
background staining

Hematoxylin and Eosin (H&E): to


determine if a fungus is hyaline or
dematiaceous

Mayers mucicarmine: to determine


mucoid capsule of C. neoformans

Fontana-Masson Stain:
demonstration of melanin or melanin
like substances in the lightly
pigmented agents of
phaeohypomycosis

Acid-Fast Stain (Kinyouns): used to


differentiate Nocardia from
Actinomyces

Acridine Orange: detects Tinea


versicolor, green fluorescent fungal
elements

Recommended incubation
time:
7 days yeast in mouth, throat and
vagina
21 days fungal pathogens in tissues
and sterile body fluids other than
blood
Up to 28 days respiratory, BM,
blood specimens and specimens in
which a dimorphic fungi is suspected

Special Tests
Hair Baiting Test differentiates T.
mentagophytes (+) from T. rubrum
(-)

Germ Tube Test initial test, isolates


yeast ; Germ tubes are elongated,
finger-like extensions from a yeast
cells ; confirmatory for the ID of C.
albicans and C. dubliniensis

Urease Production (+) alkalinisation


of media which is indicated by the
production of pink color | Christensen
Urea Agar | C. neoformans
production of urease within 4 days
help differentiate T. mentagrophytes
(urease + from T. rubrum)

L-DOPA Ferric Citrate Test rapid ID


of c. neoformans

ANTIMICROBIAL SENSITIVITY TESTING FOR


FUNGI

Commercially Available
Systems
Sensititre Yeast One Test Panel
microtitre broth dilution method,
consist to a disposable microtitre
plate, results are expressed in MIC,
excellent shelf life, for molds too.

Fungitest modified microtitre broth


breakpoint test, easy to set up and
the end points are clearly visible,
excellent shelf life, not yet evaluated
for molds

Neo-Sensitabs simple agar diffusion


method using tablets to determine
susceptibility, cheap, easy to set
up,not that reliable

Etest agar diffusion method using a


strip with a predefined concentration
gradient of the antimicrobial agent,
simple to perform and methodology
is similar to that used in bacteriology