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Thesis

English

ID: <

10670/1.bucrzw

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Diagnosis of cutaneous mycobacterioses in West Africa

Abstract

Cutaneous mycobacterioses remains prevalent in West Africa, despite the existence of treatments due to problems linked to early diagnosis. The aim of this thesis is to contribute to the improvement of their microbiological diagnosis. The results of the work of this thesis allow us to understand the epidemiology of cutaneous mycobacterioses in Burkina Faso, West Africa. This work is a first that focuses on the simultaneous microbiological diagnosis of Buruli ulcer, leprosy and cutaneous tuberculosis in Burkina Faso using innovative approaches. A review of the literature carried out on the microbiology of skin and subcutaneous ulcers in West Africa made it possible to identify the following objectives: i) to study the molecular prevalence of Mycobacterium ulcerans in clinical samples of chronic wounds in patients living in the rural areas of South West and West of Burkina Faso; ii) set up a microscopic technique for hybridization of fluorescent molecular probes for precise detection of Mycobacterium leprae in clinical skin samples; iii) used this technique to search in non-invasive samples (stool and nasal secretions) of Mycobacterium ulcerans in order to propose non-invasive methods of diagnosing leprosy in Burkina Faso, West Africa; iii) to put in place a microscopic staining technique making it possible to distinguish living, dormant or dead mycobacteria in clinical samples for monitoring treatments and to provide researchers with a tool for selecting samples eligible for attempts to culture Mycobacterium leprae. The clinical samples for the research of Mycobacterium ulcerans were made in Bobo-Dioulasso, in Djikologo (Diébougou) and in Bomborokuy (Nouna) that is 64 swabs of chronic skin wounds. As for skin samples (biopsies and skin incisions), they were carried out in the dermatology department of the “Centre Hospitalier Universitaire Souro Sanou de Bobo – Dioulasso”. Molecular tests carried out for Mycobacterium ulcerans targeting the following markers: IS2404, IS2606 and KR-B. A specific oligonucleotide probe designer on the role of Mycobacterium leprae and labeled with fluorochrome Alexa555 was used by Fluorescence microscopy - leprosy. Fluorescein diacetate, red Nile and DAPI were used for the characterization of Mycobacterium leprae according to the three stages: living, dynamic and dead. For Burkina Faso, there is a first molecular proof of the circulation of Mycobacterium ulcerans in chronic wounds of rural patients. A more specific microscopic tool made it possible to make a precise diagnosis which enabled the dermatologists of CHUSS to treat the sick. For clinicians, DDD-leprosy remains a tool for monitoring patients on anti-leprosy drugs. The researchers will find in DDD a tool to relaunch research on the culture of leprosy based on samples they have an overview of the proportion of living mycobacteria. These results confirm on the one hand a geographical extension of Buruli ulcer in West Africa and on the other hand call for a strengthening of efforts in the fight against leprosy in Burkina Faso, West Africa with the end a prospect of strengthening approaches to leprosy culture for scientists.Keywords: Cutaneous mycobacterioses / West Africa / Burkina Faso / Mycobacterium ulcerans / Mycobacterium leprae / FISH / DDD.

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