D'Avila Daniella Alchaar, Macedo Andréa Mara, Valadares Helder Magno Silva, Gontijo Eliane Dias, de Castro Ana Maria, Machado Carlos Renato, Chiari Egler, Galvão Lúcia Maria Cunha
Departamento de Parasitologia, Instituto de Ciências Biológicas, Universidade Federal de Minas Gerais, Av. Antônio Carlos, 6627 Caixa Postal 486, 31270-901, Belo Horizonte, Minas Gerais, Brazil.
J Clin Microbiol. 2009 Jun;47(6):1718-25. doi: 10.1128/JCM.01658-08. Epub 2009 Apr 8.
Our research aimed to characterize the genetic profiles of 102 Trypanosoma cruzi isolates recently obtained from 44 chronic chagasic patients from different regions of the states of Minas Gerais and Goiás in Brazil. At least two isolates were obtained from each patient at different times in order to study the parasite population dynamics during disease progression in the chronic phase. The isolates were characterized molecularly by genotyping the 3' region of the 24S alpha rRNA, the mitochondrial cytochrome oxidase subunit 2 (COII) gene, and the intergenic region of the spliced leader intergenic region (SL-IR) gene. Seventy-seven isolates were analyzed for nine microsatellite loci. The data presented here show a strong correlation between the T. cruzi lineage II (T. cruzi II) and human infection in these regions of Brazil. Interestingly, isolates from two patients were initially characterized (by rRNA genotyping) as T. cruzi I and hybrid strains, but subsequent analyses of the COII and SL-IR genes confirmed that those isolates belonged to T. cruzi III and a hybrid group, respectively. Our results confirm the risk of misclassifying T. cruzi isolates on the basis of analysis of a single molecular marker. The microsatellite profiles showed that different isolates obtained from the same patient were genetically identical and monoclonal. Exceptions were observed for T. cruzi isolates from two patients who presented differences for the SCLE11 locus and also from two other patients who showed amplification of three peaks for a microsatellite locus (TcAAAT6), implying that they were multiclonal. On the basis of the findings of the studies described here, we were not able to establish a correlation between the clinical forms of Chagas' disease and the genetic profiles of the T. cruzi isolates.
我们的研究旨在描绘最近从巴西米纳斯吉拉斯州和戈亚斯州不同地区的44名慢性恰加斯病患者身上获取的102株克氏锥虫分离株的基因图谱。为了研究慢性期疾病进展过程中寄生虫的种群动态,从每位患者身上在不同时间至少获取了两株分离株。通过对24Sα rRNA的3'区域、线粒体细胞色素氧化酶亚基2(COII)基因以及剪接前导基因间隔区(SL-IR)基因的基因分型,对这些分离株进行分子特征分析。对77株分离株分析了9个微卫星位点。此处呈现的数据表明,在巴西的这些地区,克氏锥虫II型(T. cruzi II)与人类感染之间存在很强的相关性。有趣的是,最初有两名患者的分离株(通过rRNA基因分型)被鉴定为克氏锥虫I型和杂交菌株,但随后对COII和SL-IR基因的分析证实,这些分离株分别属于克氏锥虫III型和一个杂交组。我们的结果证实了基于单一分子标记分析对克氏锥虫分离株进行错误分类的风险。微卫星图谱显示,从同一患者获得的不同分离株在基因上是相同的且为单克隆。在两名患者的克氏锥虫分离株中观察到了例外情况,这两名患者的SCLE11位点存在差异,另外两名患者的一个微卫星位点(TcAAAT6)出现了三个峰的扩增,这意味着它们是多克隆的。基于此处所述研究的结果,我们无法确定恰加斯病的临床形式与克氏锥虫分离株的基因图谱之间的相关性。