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8/12/2019 laboratory Diagnosis of Fungi
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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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THANK YOU
FOR
LISTENING.