Anna Liezle T.

Pagud MD,RMT

Cryptococcus neoformans, because
of its large polysaccharide capsule,
can be visualized by the India stain.
Organisms that possess a
polysaccharide capsule exhibit a
halo around the cell against the
black background created by the
India.

Mix the specimen with a small drop of India on a
clean glass slide.
Place a cover slip over the smear and press
gently. The preparation should be brownish, not
black.
Examine the smear microscopically for the
presence of encapsulated cells as indicated by
clear zones surrounding the cells.

Note: The India ink is ready to use. Further
dilution with water is not recommended.

Presumptive test for
Cryptococcus neoformans
Negative staining


For staining yeasts in various fluid
and secretions
Can be used to detect Histoplasma
capsulatum in bone marrow
preparations and blood smear
Can be used on sputum and respiratory
tract samples
Fastidiuous and non fastidous fungi
30 C
Dermatophytes does not grow on
higher temperature
Subcutaneous fungi - 25-30 and 37 C
H. capsulatum, Sportothrix schenckii
dimorphic
25-30 C molds Sabourauds dextrose
agar
Higher temp budding yeasts - Brain
heart infusion agar



2-4 weeks
Aspergillus fumigatus and Candida
albicans 1- 3 days
At least 3 times weekly


Blood Culture
Sabourauds dextrose agar
Dextrose , peptone, agar with ph 5.6
molds
Brain heart infusion agar
For fastidious organisms
yeast

Detection of antibodies to
specific fungal pathogens
Histoplasmosis and
Coccidiomycosis
Cryptococcis
Immunodiffusion more specific
but less sensitive
Complement fixation
Histoplasmosis and
Coccidiomycosis
Latex agglutination
Polysaccharide capsular antigens
of Cryptococcus neoformans
Enzyme Linked Immunosorbent
Assay ( ELISA )
Quantitative Enzyme
Immunoassay
HIV infected patients
Histoplasmosis
ELISA

Minimum Inhibitory Concentration
( MIC )

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