RESUMEN
Mycobacterium ulcerans infections are a public health problem in Céte d'Ivoire. The etiological diagnosis of this disease made by culture remains a big concern due to the slowness and difficulties encountered. This detection by culture of M. ulcerans represents a big interest as it allows obtaining the circulating strains for research. The purpose of this study was to determine on a routine basis in a poorly equipped laboratory, in vitro culture of M. ulcerans from exudates of skin ulcerations and from biopsy of patients with suspected Buruli ulcer. A particular attention was paid to the conditioning of the sample forwarded to the laboratory and inoculation in Lowenstein-Jensen medium supplemented with glycerol. The results of the three methods for the analysis showed 26.7, 57.4 and 17.8% positive rate respectively in the microscopy examination by nested PCR and by culture. In all the analysis, the positive rate from biopsy is higher than that obtained from exudates. The overall contamination rate by invasion of the three tubes of culture by fungi is 15.8 with 14.3 and 19.4% respectively,from exudates and biopsies. All positive samples in Ziehl-Neelsen staining and in culture were also positive by nested PCR. The nested PCR confirmed the positive strains found in culture, which were responsible for skin ulcerations. After culture, only one strain was nPCR negative. This strain was identified as Mycobacterium Gordonae. Our culture conditions showed that M. ulcerans was not the only strain identified and that other strains were present in the culture. We can conclude that the culture of M. ulcerans, in spite of the growth difficulties of the bacterium can be performed in laboratory in developing countries despite the lack of reagent and consumables. The implementation of this culture is the only way to determine sensitivity tests in vitro and in vivo in order to treat patients with Buruli ulcer.
Asunto(s)
Técnicas Bacteriológicas , ADN Bacteriano/análisis , Infecciones por Mycobacterium no Tuberculosas/diagnóstico , Mycobacterium ulcerans/crecimiento & desarrollo , Mycobacterium ulcerans/aislamiento & purificación , Biopsia , Côte d'Ivoire , Medios de Cultivo , Exudados y Transudados/microbiología , Humanos , Infecciones por Mycobacterium no Tuberculosas/microbiología , Mycobacterium ulcerans/genética , Reacción en Cadena de la Polimerasa , Úlcera Cutánea/microbiologíaRESUMEN
The Côte-d'Ivoire Pasteur Institute unit of Tuberculous and Nontuberculous Mycobacteria carried out 600 smears stained by Ziehl-Neelsen in 200 immigrant candidates for the U.S.A. The sputa of 44 of them were put in culture on Lowenstein-Jensen medium. Eight (4%) candidates had active pulmonary tuberculosis among whom 5 had smear negative sputum. The pulmonary tuberculosis active detection performed on target population with accessible and sensitive tool can contribute to strengthen the fight against tuberculosis in endemic areas.