Dr. F.

Kechia Generalities on Diagnosis

Diagnosis
Like all microbial infection, the diagnosis of mycosis is based on epidemiological data, clinical data, and laboratory data.

Diagnosis
Laboratory (Biological) examination comprises
direct examination of pathological samples in wet mount after coloration or not, or after softening, the culture and identification of the fungus, the detection of a specific immunological reaction, humoral and / or cellular and the detection of circulating antigens. Recently molecular detection for certain mycoses has been made possible.

Diagnosis
Clinical diagnosis- the different clinical signs of mycoses can furnish a presumptive diagnosis of a fungal infection. Superficial and sub-cutanous mycoses can produce characteristic lesions but can equally present some aspect that are non specific and atypical (particularly in the case of previous treatment with topical corticoids)

Diagnosis
In deep mycoses, the clinical signs are generally not specific but constitute some elements of orientation of high value to shade light on the underlying situation, geographical or ethnical , the viscera attained.
Certain complementary examinations can be of great help (detection cryptococcal antigen, scanning in hepato-splenic candidoses.

Diagnosis
Laboratory diagnosis Samples, when possible, permit to affirm the diagnosis by
direct examination, culture, histology, immunology, animal inoculation molecular technique.

Diagnosis
Sample diagnosis of a mycosis depends much on the manner in which the pathological sample is taken or collected. A good diagnosis can not be done on a badly taken sample. Samples should be collected from the right part of the lesion, in sufficient quantity and in good conditions.

Diagnosis
Direct examination (DE) all pathological materials ( CSF, expectoration, BAL, swabs, stool?, urine, gastric washings, biopsies etc) submitted to the laboratory for mycological examination must be examined by direct microscopy. DE can be with or without coloration of the material. DE has as objective to observe the fungal responsible and to determine the qnty of the fungal elements (FE) in the sample.

Diagnosis
Coloration with lactophenol cotton blue is convenient for all sample types; but coloration with Gram is conveneint for most types of materials and is positive for all fungi in good physiological state. Samples such as hair, nail should be digested with KOH or chlorolactphenol to make the FEs visible. This of interest during the search for dermatophytes.

Diagnosis
Wet mounts (pus, expectoration) should be done in Gram, Methylene blue, Giemsa, meanwhile anatomopathological sections should be colored with Gomori-GrocottHotchkiss-Mc Manus. Histopathological occupy first place in the study of sub-cutanous and deep mycoses: inflammatory reactions and modification of tissues often have particular aspects which orients to the research.

Diagnosis
Mycotic inflammation is often characterized by sub acute or chronic granulomatous reaction to which is associated suppuration and fibrosis.

Diagnosis
Planting and isolation Culture of fungi is made possible on classical culture media (Sab, Malt extract) but sometimes difficult (Histoplasma) or impossible (Rhinosporidium, Pneumocystis). Certain fungi demand special growth factors (vitamins for Trichophyton ochraceum) or special culture media ( blood medium for the yeast phase of dimorphic fungi. Note use of antibiotics in fungal culture media.

Diagnosis
Fungi of superficial develop at ordinary temperature while those of deep mycoses grow at 37C this differenciates a number saprophytic fungi which do not grow at this temperature 37C.

Diagnosis
Yeast develop in 24-48h, moulds 2-4days, the dermatophytes and most pathogenic fungi 615days

Diagnosis
Identification -macroscopic morphology -microscopic morphology -physiology of fungi on specific media permit the identification of the genus and species

Diagnosis
Morphological criteria - capsule (C. neoformans) - germinative tube pseudo-filament (C. albicans) - arthrospore (Trichosporon and Geotrichum) - chlamydospore (C. albicans) - unipolar bud (Malassezia) Conidia head ( Aspergillus, penicillium)

Diagnosis
Physiological criteria - Zymogram - Auxanogram - Enzymatic activity:-Urease, phenoloxydase.

Diagnosis
Immunological criteria - these interest serotyping eg A or B of C. albicans and A, B, C OR D of C. neoformans.

Diagnosis
Indirect Diagnostic methods - Cutanous activity: intradermo-reactions exploit hypersensitivity - Serological reaction: - demonstrate presence of antibodies 1-Precipitation rxn, 2immunofluorescence 3-ELISA -research of circulating antigens (Aspergillosis, cryptococcosis histoplasmosis)