1. Principles of LDG of SFI: • Hair • Nail • Skin • Mucosal • Eye • Ear 1. Principles of LDG of SFI: • Hair • Nail • Skin • Mucosal • Eye • Ear The goal of teaching: 2. Principles of LDg of IFI: agglutination, precipitation, ELISA, PCR, PH invasive aspergillosis (IA) invasive candidiasis (IC) zigomycosis Pneumocystis pneumonia 2. Principles of LDg of IFI: agglutination, precipitation, ELISA, PCR, PH invasive aspergillosis (IA) invasive candidiasis (IC) zigomycosis Pneumocystis pneumonia 3. Methods for fungal identification Test methods Fungal sensitivity to AM • Diffusion • Dilution • Etest 3. Methods for fungal identification Test methods Fungal sensitivity to AM • Diffusion • Dilution • Etest 4. Epidemiology of FI • Risk groups of patients: Endemic FI (biphasic and subcutaneous mycoses) • Mycoallergens and mycoallergosis • Mycotoxins and mycotoxicosis 4. Epidemiology of FI • Risk groups of patients: Endemic FI (biphasic and subcutaneous mycoses) • Mycoallergens and mycoallergosis • Mycotoxins and mycotoxicosis
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1. Principles of LDG of SFI:• Hair• Nail• Skin• Mucosal• Eye• Ear
1. Principles of LDG of SFI:• Hair• Nail• Skin• Mucosal• Eye• Ear
The goal of teaching: 2. Principles of LDg of IFI:
3. Methods for fungal identification Test methods Fungal sensitivity to AM
• Diffusion• Dilution• Etest
3. Methods for fungal identification Test methods Fungal sensitivity to AM
• Diffusion• Dilution• Etest
4. Epidemiology of FI
• Risk groups of patients:
Endemic FI (biphasic and subcutaneous mycoses)
• Mycoallergens and mycoallergosis
• Mycotoxins and mycotoxicosis
4. Epidemiology of FI
• Risk groups of patients:
Endemic FI (biphasic and subcutaneous mycoses)
• Mycoallergens and mycoallergosis
• Mycotoxins and mycotoxicosis
LDg and localization of FI
LDg of FI: METHODS
Classical / traditional / mycological (conventional) methods:• DMP / PH• CULTURE (isolation) the "golden standard"
Most fungi are cultivated on nutritious media (NM), except
Pneumocystis, Rinosporidium ... - The problem - a low sensitivity (IFI) - analysis are long-lasting
Classical / traditional / mycological (conventional) methods:• DMP / PH• CULTURE (isolation) the "golden standard"
Most fungi are cultivated on nutritious media (NM), except
Pneumocystis, Rinosporidium ... - The problem - a low sensitivity (IFI) - analysis are long-lasting
- Immunodiagnosis (method of choice for IFI and endemic FI)• Detection of Ag (agglutination, diffusion, ELISA)• Ab detection (agglutination, diffusion, ELISA)
The problem - interpretation of the results
- Molecular diagnosis (rapid and sensitive)• DNA detection (PCR)
The problem - contamination standardization
- Immunodiagnosis (method of choice for IFI and endemic FI)• Detection of Ag (agglutination, diffusion, ELISA)• Ab detection (agglutination, diffusion, ELISA)
The problem - interpretation of the results
- Molecular diagnosis (rapid and sensitive)• DNA detection (PCR)
The problem - contamination standardization
Processing
Classic / Traditional (conventional) method
• 4-7/14-21 day (dermatophytes)
• Low sensitivity, slow
Proven FI
Classic / Traditional (conventional) method
• 4-7/14-21 day (dermatophytes)
• Low sensitivity, slow
Proven FI
Immunological methodsMolecular methods
• ~ up to 24 hours
• Highsensitive, fast
Probable FI
Immunological methodsMolecular methods
• ~ up to 24 hours
• Highsensitive, fast
Probable FI
LDg of FI - METHODS
Request for analysis and transport
Antifungal susceptibility testing
Patient
Sampling
Sputum
Symptoms and signs of infectionUrine
Blood
Feces
Tissue
SwabHypothesis
Sample for anaerobes
Preparation Culture Serology Molecular methods
QuantitationIdentification
Agglutination Hybridisation
Diagnosis
Therapy Patient without symptoms
Superficial fungal infections (SFI) • Proper selection and sampling:
- prior to initiation of Th - the right place - in sufficient quantity - asepsis!
• Proper selection and sampling:
- prior to initiation of Th - the right place - in sufficient quantity - asepsis!
Direct microscopic examination (DME): A quick, inexpensive method
• Only reveales fungal elements• It is not possible to identify
Direct microscopic examination (DME): A quick, inexpensive method
• Only reveales fungal elements• It is not possible to identify
• Isolation of fungi: slow method, but the "golden standard“• Identification of fungi, quantization, typing, susceptibility
testing...
• Isolation of fungi: slow method, but the "golden standard“• Identification of fungi, quantization, typing, susceptibility
testing...
Wood-ova lampa – neke vrste dermatofita fluorescirajuHair - proper sampling
Hair: Wood’s lamp
Microsporia - some strains of Microsporum spp. have the ability to autofluorescence under UV light
Microsporia - some strains of Microsporum spp. have the ability to autofluorescence under UV light
Hair – proper sampling
FavusFavus
139Instruments for sampling in suspected SFI
(nail, hair)
Psoriasis Onychomycosis Psoriasis Onychomycosis
Nail – proper sampling
Nail – proper sampling
Finger-nail scrapings
Finger-nail scrapings
• DMP• CULTURE
• DMP• CULTURE
Nail – proper sampling
Skin – proper sampling
Skin squama Skin squama • DMP• CULTURE
• DMP• CULTURE
“Screeping” cornea of the eye - the proper
sampling
KeratomycosisKeratomycosis The finding of hyphae in corneal scraping (KOH)
The finding of hyphae in corneal scraping (KOH)
Ear – proper sampling
Isolated A. fumigatusIsolated A. fumigatus
Isolated C. parapsilosisIsolated C. parapsilosis
The material is taken using a smear The material is taken using a smear