Calcagno A M, Niño-Vega G, San-Blas F, San-Blas G
Instituto Venezolano de Investigaciones Cientificas, Centro de Microbiología y Biología Celular, Laboratorio de Micrología, Caracas, Venezuela.
J Clin Microbiol. 1998 Jun;36(6):1733-6. doi: 10.1128/JCM.36.6.1733-1736.1998.
Randomly amplified polymorphic DNA (RAPD) analysis of 33 Paracoccidioides brasiliensis strains from Argentina, Brazil, Colombia, Peru, and Venezuela produced reproducible amplification products which were sufficiently polymorphic to allow differentiation of the strains. Types generated with five primers (OPG 03, OPG 05, OPG 14, OPG 16, and OPG 18) resulted in a high discriminatory index (0.956). The discriminatory index was slightly reduced (0.940) when only two primers (OPG 3 and OPG 14) were used. A dendrogram based on these results showed a high degree of similarity among the strains, and genetic differences were expressed in clusters related to geographical regions but not to pathological features of the disease. With a few exceptions, strains were sorted into five groups by geographical origin as follows: group I, Venezuelan strains; group II, Brazilian strains; group III, Peruvian strains; group IV, Colombian strains; and group V, Argentinian strains. The group containing the most disparate strains was group V (discriminatory index, 0.633); the discriminatory index for the other four groups was 0.824. The use of primer OPG 18 by itself was sufficient to discriminate species specificity, and the use of primer OPG 14 by itself was sufficient to discriminate among the geographical locations of the strains in the sample. This method may be helpful for epidemiological studies of P. brasiliensis.
对来自阿根廷、巴西、哥伦比亚、秘鲁和委内瑞拉的33株巴西副球孢子菌进行随机扩增多态性DNA(RAPD)分析,产生了可重复的扩增产物,这些产物具有足够的多态性,能够区分这些菌株。用5种引物(OPG 03、OPG 05、OPG 14、OPG 16和OPG 18)产生的类型具有较高的鉴别指数(0.956)。当仅使用两种引物(OPG 3和OPG 14)时,鉴别指数略有降低(0.940)。基于这些结果的树状图显示菌株之间具有高度相似性,遗传差异在与地理区域相关的簇中表现出来,而与疾病的病理特征无关。除少数例外,菌株按地理来源分为五组,如下:第一组,委内瑞拉菌株;第二组,巴西菌株;第三组,秘鲁菌株;第四组,哥伦比亚菌株;第五组,阿根廷菌株。包含最不同菌株的组是第五组(鉴别指数,0.633);其他四组的鉴别指数为0.824。单独使用引物OPG 18足以区分物种特异性,单独使用引物OPG 14足以区分样品中菌株的地理位置。该方法可能有助于巴西副球孢子菌的流行病学研究。