Campos R F, Gonçalves M S, dos Reis E A, dos Reis M G, Andrade S G
Laboratório de Doença de Chagas Experimental e Laboratório de Patologia e Biologia Molecular, Centro de Pesquisas Gonçalo Moniz-Fiocruz, BA, Brasil.
Mem Inst Oswaldo Cruz. 1999 Jan-Feb;94(1):23-9. doi: 10.1590/s0074-02761999000100009.
Molecular characterization of one stable strain of Trypanosoma cruzi, the 21 SF, representative of the pattern of strains isolated from the endemic area of São Felipe, State of Bahia, Brazil, maintained for 15 years in laboratory by serial passages in mice and classified as biodeme Type II and zymodeme 2 has been investigated. The kinetoplast DNA (kDNA) of parental strain, 5 clones and 14 subclones were analyzed. Schizodeme was established by comparative study of the fragments obtained from digestion of the 330-bp fragments amplified by polymerase chain reaction (PCR) from the variable regions of the minicircles, and digested by restriction endonucleases Rsa I and Hinf I. Our results show a high percentual of similarity between the restriction fragment length polymorphism (RFLP) for the parental strain and its clones and among these individual clones and their subclones at a level of 80 to 100%. This homology indicates a predominance of the same "principal clone" in the 21SF strain and confirms the homogeneity previously observed at biological and isozymic analysis. These results suggest the possibility that the T. cruzi strains with similar biological and isoenzymic patterns, circulating in this endemic area, are representative of one dominant clone. The presence of "principal clones" could be responsible for a predominant tropism of the parasites for specific organs and tissues and this could contribute to the pattern of clinico-pathological manifestations of Chagas's disease in one geographical area.
对一种克氏锥虫稳定菌株21 SF进行了分子特征分析。该菌株是从巴西巴伊亚州圣费利佩流行地区分离出的菌株模式的代表,通过在小鼠体内连续传代在实验室中保存了15年,被分类为生物型II和酶型2。对亲代菌株、5个克隆和14个子克隆的动基体DNA(kDNA)进行了分析。通过比较从微小环可变区经聚合酶链反应(PCR)扩增得到的330 bp片段,用限制性内切酶Rsa I和Hinf I消化后获得的片段,确定了裂殖体型。我们的结果显示,亲代菌株及其克隆以及这些单个克隆与其子克隆之间的限制性片段长度多态性(RFLP)相似性百分比很高,在80%至100%之间。这种同源性表明在21SF菌株中同一“主要克隆”占主导地位,并证实了先前在生物学和同工酶分析中观察到的同质性。这些结果表明,在该流行地区传播的具有相似生物学和同工酶模式的克氏锥虫菌株可能代表一个优势克隆。“主要克隆”的存在可能是寄生虫对特定器官和组织具有主要嗜性的原因,这可能导致恰加斯病在一个地理区域的临床病理表现模式。