Diagnosis of Fungal Infections (1)[1]

GROUP 5
MEMBERS
1. ASHU NARDINE
2. QUEEN CHIDIRA CHIJOKE
3. ASSAH ELVINE
4. SAME SHARON
5. AYUKETA-NCHI
6. MESODE MOLLY
7. EkOLE WILLIAM
8. SOLKEM EMMANUELLE CHELA
9. TAKUMBI PETERKINS
DIAGNOSIS OF FUNGAL INFECTIONS
OUTLINE
 Clinical presentation and History
 Symptoms and clinical signs of fungal
infections
 Laboratory technique
 Microscopy; KOH staining, gram staining
 Culturing fungi; Saboaraud agar, other
selective media
 Molecular methods;PCR, DNA Sequencing
 Histopathology
Fungal identification in tissue samples
CLINICAL PRESENTATION AND HISTORY OF
FUNGAL INFECTIONS
History
• They are caused by fungi that exists in
our every day environment.
• Hippocrates wrote on sores in the mouth
in 500BC which modern mycologist
identified a s thrust.
• More than 1000 years after ringworm
infections was present in the skin and
hair.
• In the modern era (1842-1844) David Grubby
• The work of Louis pasteur and Agostino
Bassi an Italian entomologist showed that
fungal infections were and still are the
cause of economic problems in agriculture
and related industries.
 Clinical presentation
 Skin lesions
 Weight loss
Swollen lymph node Discharge or pus
 Fatigue

SYMPTOMS AND CLINICAL S I GN S OF FUNGI
INFECTION
 Common symptoms
 Skin lesions or rashes
 Itching, burning or redness
 Swelling, infl ammation or redness
 Discharge
 Fever, fatigue, headache, muscle or
joint pain
 Clinical Signs
1) For s kin a nd m ucus membrane
 Youpapules (raised ,solid, lesions)
 vesicles(fluid,filled, lessions)
 ulcers(open sores)
 Erythema(redness)
 Edema(swelling)
 Nails
 nails separation
 subungual debris(debris under the nail)
 Hair
 Alopecia(hair loss)
 scalp lessions
 broken hair
 Eye
oconjunctivities(red, itchy, eye)
Laboratory technique
o There are several laboratory techniques used
to identify fungal infections.
o These include direct microscopic examination
of clinical samples, culture, antigen testing,
serological testing, and PCR are also used for
diagnosis of fungal infections.
 Direct Microscopic
Examination
• KOH (potassium hydroxide) preparation: This

is a simple and rapid method for direct
examination of fungal elements in clinical
samples.
• Calcofluor white stain: This stain binds to
fungal elements in a sample and fl uoresces
 Culture
Fungal culture: This involves growing fungi
from a clinical sample on a nutrient mFungal
o identify the species.
 Serological Testi ng
Detection of antibodies against fungal
antigens in blood samples.
 PCR (Polymerase Chain Reaction)
A molecular biology technique used to amplify

specifi c DNA sequences from fungal genomes.
Microscopy; KOH staining, gram staining
 Microscopy: KOH preparation
KOH serves a s an enzymatic agent that breaks down
debris in a specimen to view hyphae or any yeast
cell.
 Place the specimen on a clean slide

 Add a drop of about 20% KOH(concentration depends
on the specimen) and smear.
 Place a cover glass and press gentle to get rid
of air bubbles.
 Blot excess solution from the finished slide

preparation.
 Place slide on the microscope stage and start

with low power(10x) to make cells more visible.
 Examine for fungal structures. Presence of

hyphae or yeast suggest fungus.
 Gram staining
Here, we prepare specimen smear on slide
 Fix with heat

 Place drops of crystal violet (primary stain)for about
1minute, rinse with water
 Iodine(act a s a mordant) is applied in the slide for about
1minute and rinsed with water
 Acetone (decolorizer) is then put on the slide for about
2-4seconds and rinsed immediately
 Safranin(counterstain) is also applied on the slide for
about 1min and rinsed
 Examine slide for the presence of yeast, hyphae
and spores stained indicating fungi infection .
CULTURING FUNGI SABOARAUD AGAR, OTHER
SELECTIVE MEDIA
Culturing fungi; Sabouraud agar, other selective

media*
-Culture media
-Selective media Sabouraud Agar
 Composition
•Peptones: Provide nitrogen and carbon
sources
•Dextrose: Sugar that supports fungal growth
•Agar: Solidifying agent
 Usage
•ideal for the cultivation of dermatophytes.
•can be supplemented with antibiotics like
chloramphenicol to inhibit bacterial growth
Other selective media
Potato Dextrose Agar
 Czapek Dox Agar
 Cornmeal Agar
Malt Extract Agar Culturing process
•Preparation
• Inoculation
• Incubation
•O bs ervation Identification
•Use of microscope Safety considerations
• Work in a biosafety cabinet

• Wear PPE
Molecular methods;PCR, DNA Sequencing
Molecular diagnosis of fungal by PCR

PCR (Polymerase Chain Reaction)
Advantages:
 Sensitivity
 Specificity
 -fast Result-Non-invasiv
 Types of PCR for fungal diagnosis:

 Conventional PCR: Detects specifi c DNA
Real-time PCR (PCR): Quantifies fungal DNA, helping
monitor treatment response.
Nested PCR: Increases sensitivity for detecting rare or
degraded DNA.
Multiplex PCR: Simultaneously detects multiple
fungal species.
 Clinical applications:
nvasive aspergillosis diagnosis
 Candidemia detection
 Cryptococcal meningitis diagnosis
Pneumocystis pneumonia detection
Sample type to be collected include; Blood,Tissue
biopsies,Cerebrospinal fluid(CSF), Respiratory
secretions,Skin.
Histopathology
Histopathology of fungal infections involves the
examination of tissue samples under a microscope
to identify fungal elements.
 HISTOPATHOLOGICAL FEATURES
GENERAL FEATURES
Inflammation(acute or chronic)
 Tissue necrosis
Fungal elements (hyphae, yeast, spores)
 SPECIFIC FEATURES
YEAST e.g Candida, histoplasma,

crytococcus
 Budding yeast cells
 Pseudohyphae
 Blastoconidia
 Molds e.g Aspergillus
 Hyphae
 Spores
 Conidiophores
FUNGAL IDENTIFICATION IN TISSUE SAMPLES
Fungal identification in tissue samples is a
step in diagnosing fungal infections.
Several methods can be used to identify
fungi in tissue samples, including:
• Microscopic examination:
This involves examining tissue sections
under a microscope to look for fungal
elements, such a s hyphae, yeast cells, or
spores.
• Culture: This involves growing fungi from
tissue samples in a laboratory to identify
the species.
• Molecular tests: These involve the use of

DNA-based techniques
Limitations of Fungal Identification
 Contamination: Tissue samples can be

contaminated with fungi from the
environment or from the laboratory, leading to
false- positive results.
 Limited sensitivity: Some fungal infections may
not be detectable by current methods,
leading to false-negative results.
 Diffi culty in disti nguishing between species: Some
fungal species can be diffi cult to distinguish
from one another, leading to incorrect

Diagnosis of Fungal Infections (1)[1]

Uploaded by

Copyright:

Available Formats

Diagnosis of Fungal Infections (1)[1]

Uploaded by

Document Information

Copyright

Available Formats

Share this document

Share or Embed Document

Sharing Options

Did you find this document useful?

Is this content inappropriate?

Copyright:

Available Formats

Diagnosis of Fungal Infections (1)[1]

Uploaded by

Copyright:

Available Formats

GROUP 5

• KOH (potassium hydroxide) preparation: This

 PCR (Polymerase Chain Reaction)

A molecular biology technique used to amplify

 Place the specimen on a clean slide

 Blot excess solution from the finished slide

 Place slide on the microscope stage and start

 Examine for fungal structures. Presence of

Here, we prepare specimen smear on slide

 Fix with heat

Culturing fungi; Sabouraud agar, other selective

Other selective media

Potato Dextrose Agar

 Czapek Dox Agar

• Work in a biosafety cabinet

Molecular diagnosis of fungal by PCR

 Types of PCR for fungal diagnosis:

YEAST e.g Candida, histoplasma,

• Molecular tests: These involve the use of

 Contamination: Tissue samples can be

You might also like