Coulibaly B, Coulibaly-N'Golo M-D G, Ekaza E, Aka N, N'Guessan K R, Baudryard A, Assandé J-M, Trébissou N, Guédé-Guina F, Dosso M
Unité des mycobactéies tuberculeuses et atypiques, département de bactéiologie-virologie, institut Pasteur de Céte-d'Ivoire, BP 490 Abidjan 01, Céte-d'lvoire.
Bull Soc Pathol Exot. 2010 Feb;103(1):2-7. doi: 10.1007/s13149-009-0002-y.
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.
溃疡分枝杆菌感染是科特迪瓦的一个公共卫生问题。由于培养过程缓慢且存在困难，通过培养进行该疾病的病因诊断仍是一个重大问题。通过培养检测溃疡分枝杆菌具有重要意义，因为它能获取用于研究的流行菌株。本研究的目的是在设备简陋的实验室中，常规地从皮肤溃疡渗出物和疑似布鲁里溃疡患者的活检组织中进行溃疡分枝杆菌的体外培养。特别关注了送往实验室的样本处理以及在添加甘油的罗-琴培养基中的接种。三种分析方法的结果显示，巢式PCR显微镜检查、普通PCR和培养的阳性率分别为26.7%、57.4%和17.8%。在所有分析中，活检组织的阳性率高于渗出物的阳性率。三种培养管被真菌污染的总体污染率为15.8%，渗出物和活检组织的污染率分别为14.3%和19.4%。萋-尼染色和培养中的所有阳性样本通过巢式PCR也呈阳性。巢式PCR证实了培养中发现的导致皮肤溃疡的阳性菌株。培养后，只有一株菌株巢式PCR呈阴性，该菌株被鉴定为戈登分枝杆菌。我们的培养条件表明，溃疡分枝杆菌不是唯一被鉴定出的菌株，培养中还存在其他菌株。我们可以得出结论，尽管该细菌生长困难，但在发展中国家，尽管缺乏试剂和耗材，溃疡分枝杆菌的培养仍可在实验室中进行。实施这种培养是确定体外和体内敏感性试验以治疗布鲁里溃疡患者的唯一途径。
