Mbwana J, Bölin I, Lyamuya E, Mhalu F, Lagergård T
Department of Medical Microbiology and Immunology, The Sahlgrenska Academy, Göteborg University, Sweden.
J Clin Microbiol. 2006 Jan;44(1):132-7. doi: 10.1128/JCM.44.1.132-137.2006.
The technique of random amplified polymorphic DNA (RAPD) was adapted and optimized to study Haemophilus ducreyi isolates. A panel of 43 strains isolated from chancroid patients from different countries in Africa, Europe, North America, and Asia were characterized. The strains were also studied with respect to lipooligosaccharide (LOS) migration and immunoblotting patterns and the presence of cytolethal distending toxin genes. The RAPD method with the OPJ20 primer generated nine banding patterns (1 to 9). The majority of the isolates were clustered into two major profiles, 14 and 13 strains into profiles 1 and 2, respectively, and just a few strains revealed patterns 3 and 4. The isolates from Thailand were exceptional in that they showed greater diversity and were represented by six different RAPD patterns, i.e., patterns 3 and 5 to 9. The LOS migration and immunoblotting analyses revealed two different patterns, which indicated long and short forms of LOS; the former was found in 20/23 tested strains. Two strains that expressed the short form of LOS were grouped into RAPD pattern 4. The absence of cdtABC genes was observed in only 4/23 strains, and three of these isolates were assigned to RAPD pattern 4. Our results showed limited genotypic and phenotypic variations among H. ducreyi strains, as supported by the conserved RAPD and LOS profiles shared by the majority of the studied strains. However, the RAPD method identified differences between strains, including those from different geographic areas, which indicate the potential of RAPD as an epidemiological tool for the typing of H. ducreyi isolates in countries where chancroid is endemic.
随机扩增多态性DNA(RAPD)技术被应用并优化用于研究杜克雷嗜血杆菌分离株。对从非洲、欧洲、北美和亚洲不同国家的软下疳患者中分离出的43株菌株进行了特征分析。还对这些菌株的脂寡糖(LOS)迁移和免疫印迹模式以及细胞致死性膨胀毒素基因的存在情况进行了研究。使用OPJ20引物的RAPD方法产生了9种条带模式(1至9)。大多数分离株聚为两个主要类型,分别有14株和13株属于类型1和类型2,只有少数菌株呈现模式3和模式4。来自泰国的分离株很特别,它们表现出更大的多样性,由6种不同的RAPD模式代表,即模式3和模式5至9。LOS迁移和免疫印迹分析揭示了两种不同模式,表明LOS有长型和短型;在23株测试菌株中有20株发现为前者。表达LOS短型的两株菌株被归为RAPD模式4。仅在4/23株菌株中未观察到cdtABC基因,其中3株分离株被归为RAPD模式4。我们的结果显示杜克雷嗜血杆菌菌株之间的基因型和表型变异有限,这得到了大多数研究菌株共有的保守RAPD和LOS图谱的支持。然而,RAPD方法识别出了菌株之间的差异,包括来自不同地理区域的菌株,这表明RAPD作为一种流行病学工具在软下疳流行国家用于杜克雷嗜血杆菌分离株分型的潜力。