Molecular Approaches in the Diagnosis of Dermatophytosis

Dermatophytosis is one of the most common infectious diseases in the world and can be caused by several dermatophyte species. These species are closely related in genetic structure in spite of different phenotypic and ecological features. The morphological similarity, variability, and polymorphism of dermatophytes have meant that species identification for dermatophytes is time consuming and requires a significant degree of knowledge and technological expertise. Molecular biology-based techniques have solved problems concerning the morphology-based identification of dermatophytes and have improved our knowledge on the epidemiology of dermatophytosis. Further development of molecular diagnosis of dermatophytosis requires the investigation of additional molecular markers for diagnostic tools targeting multiple loci as well as the improvement of techniques.     

Diagnosis of Dermatophytosis

Dermatophytoses are infections of the skin, hair or nails caused by dermatophytes. Dermatophytes can induce typical diagnostic clinical lesions (tinea), but can also mimic other dermatoses. Therefore, physicians need to be familiar with the whole spectrum of tinea and must constantly be mindful of possible dermatophytosis. An examination with Wood’s light can be helpful. In superficial lesions, the demonstration of dermatophytes may be achieved by dermatoscopy or laser scanning. An essential step of the following diagnostic procedure is the skilful collection of samples for the proof of fungi. Microscopy of KOH mounts or equivalent preparations produced from infected material and histology are approved methods. The necessary identification of dermatophytes on species level can finally be accomplished by conventional methods based on cultures or by new techniques based on molecular differentiation or MALDI-TOF analysis. These modern methods are clearly on the increase and can considerably improve and accelerate dermatophyte identification.     

Conventional Methods for the Diagnosis of Dermatophytosis

Dermatophytes are keratinolytic fungi responsible for a large variety of diseases that can affect glabrous skin, nails and hair. In many cases, the diagnosis is not clinically obvious, and mycological analysis is required. This includes both direct microscopic examination and cultures. First of all, clinical specimens have to be sampled according to localization and characteristics of the lesions. Direct microscopic examination is usually performed using clearing reagents (KOH or Amman's chloral-lactophenol), but its sensitivity may be greatly enhanced by the use of stains or fluorochromes such as Congo red or Calcofluor white. Histological analysis is an efficient method, but it is constraining for the patients and, as direct examination, it does not allow precise identification of the pathogen. Cultures are therefore needed, and specific culture media may be used to overcome the growth of rapidly growing contaminating moulds which may hamper the recovery of dermatophytes. Identification at the species level which may be useful to initiate an appropriate treatment or for setting prophylactic measures, relies on macroscopic and microscopic morphology. Subcultures on culture media which stimulate conidiation and, for some species, the production of pigments, are often necessary. Additionally, in case of atypical isolates, some biochemical or physiological tests may be performed such as the search for urease activity or the in vitro hair perforation test. However, their contribution to species identification is rather limited, and progress is still needed for the development of biochemical or immunological tests allowing an accurate identification at the species level, pending for the availability of molecular biology-based kits.     

Plant Molecular Biology

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Plant Molecular Biology     

Bedside Diagnosis for Disseminated Deep Dermatophytosis: a Case Series Study

Mycopathologia - Deep cutaneous fungal infections including deep dermatophytosis are responsible for significant morbidity and mortality, especially in immunocompromised patients. Variable and...     

Immunology of Dermatophytosis

The immune response to infection by dermatophytes ranges from a non-specific host mechanism to a humoral and cell-mediated immune response. The currently accepted view is that a cell-mediated immune response is responsible for the control of dermatophytosis. Indeed, some individuals develop a chronic or recurrent infection mediated by the suppression of a cell-mediated immune response. The immune response to Trichophyton is unusual in that this fungus can elicit both immediate hypersensitivity (IH) and delayed-type hypersensitivity (DTH) in different individuals when they are submitted to a skin test reaction. Understanding the nature and function of the immune response to dermatophytes is an exciting challenge that might lead to novel approaches in the treatment and immunological prophylaxis of dermatophytosis.     

Pathogenesis of Dermatophytosis

Despite the superficial localization of most dermatophytosis, host-fungus relationship in these infections is complex and still poorly elucidated. Though many efforts have been accomplished to characterize secreted dermatophytic proteases at the molecular level, only punctual insights have been afforded into other aspects of the pathogenesis of dermatophytosis, such as fungal adhesion, regulation of gene expression during the infection process, and immunomodulation by fungal factors. However, new genetic tools were recently developed, allowing a more rapid and high-throughput functional investigation of dermatophyte genes and the identification of new putative virulence factors. In addition, sophisticated in vitro infection models are now used and will open the way to a more comprehensive view of the interactions between these fungi and host epidermal cells, especially keratinocytes.     

真菌果胶酶的分子生物学研究进展

近10年来国外在果胶酶分子生物学研究上取得了重大进展,从11个属的真菌克隆了50个以上的基因并测序。对果胶酶基因的结构、功能、调控、录译后加工等方面进行了深入探讨。已克隆的果胶酶基因以多聚半乳糖醛酸酶(PG)基因和果胶裂解酶(PL)基因为主,也有果胶酯酶(PE)基因和鼠李半乳糖醛酸酶(RHG)基因,大多有内含子。前体蛋白一般有N信号肽和糖基化位点。果胶酶一般受果胶、低浓度的(01%)D半乳糖醛酸等诱导,而受较高浓度(1%)的半乳糖醛酸、抗体、某些抗菌素抑制。