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Anna Liezle T. Pagud MD,RMT

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Microscopic

Examination

Culture Isolation and

Identification

Serologic Test

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10-20 KOH

Lactophenol cotton blue

Methylene blue

Gram staining

Giemsa

and rights stain

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ryptococcus 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.

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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, notblack.

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.

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Presumptive test for

Cryptococcus neoformans

Negative staining

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Gram stain

For staining yeasts in various fluid

and secretions

Giemsa

and rights

Can be used to detect Histoplasma

capsulatum in bone marrow

preparations and blood smear

Papanicolaou stain

Can be used on sputum and respiratory

tract samples

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Fastidiuous and non fastidous fungi30 C

Dermatophytes does not grow on

higher temperatureSubcutaneous fungi - 25-30 and 37 C

H. capsulatum, Sportothrix schenckii

dimorphic25-30 C molds Sabourauds dextrose

agar

Higher temp budding yeasts - Brain

heart infusion agar

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2-4 weeks

Aspergillus fumigatus and Candida

albicans 1- 3 days

At least 3 times weekly

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Blood Culture

Sabouraudsdextrose agarDextrose , peptone, agar with ph 5.6

molds

Brain heart infusion agarFor fastidious organisms

yeast

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Detection of antibodies to

specific fungal pathogens

Histoplasmosis andCoccidiomycosis

Cryptococcis

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Immunodiffusion more specificbut less sensitive

Complement fixation

Histoplasmosis andCoccidiomycosis

Latex agglutination

Polysaccharide capsular antigensof Cryptococcus neoformans

Enzyme Linked Immunosorbent

Assa ELISA

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Quantitative Enzyme

Immunoassay

HIV infected patientsHistoplasmosis

ELISA

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Minimum Inhibitory Concentration

( MIC )

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