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在位于巴西里约热内卢的伊瓦尔多·查加斯国家传染病研究所(巴西奥斯瓦尔多·克鲁兹基金会)随访的慢性恰加斯病患者分离株中发现的克氏锥虫I基因型。

Trypanosoma cruzi I genotype among isolates from patients with chronic Chagas disease followed at the Evandro Chagas National Institute of Infectious Diseases (FIOCRUZ, Brazil).

作者信息

Oliveira Tatiana da Silva Fonseca de, Santos Barbara Neves Dos, Galdino Tainah Silva, Hasslocher-Moreno Alejandro Marcel, Bastos Otilio Machado Pereira, Sousa Maria Auxiliadora de

机构信息

Coleção de Tripanossomatídeos, Instituto Oswaldo Cruz, Rio de Janeiro, RJ, Brasil.

Laboratório Interdisciplinar de Pesquisas Médicas, Instituto Oswaldo Cruz, Rio de Janeiro, RJ, Brasil.

出版信息

Rev Soc Bras Med Trop. 2017 Jan-Feb;50(1):35-43. doi: 10.1590/0037-8682-0406-2016.

Abstract

INTRODUCTION

: Trypanosoma cruzi is the etiologic agent of Chagas disease in humans, mainly in Latin America. Trypanosome stocks were isolated by hemoculture from patients followed at Evandro Chagas National Institute of Infectious Diseases (FIOCRUZ) and studied using different approaches.

METHODS

: For species and genotype identification, the stocks were analyzed by parasitological techniques, polymerase chain reaction assays targeted to specific DNA sequences, isoenzyme patterns, besides sequencing of a polymorphic locus of TcSC5D gene (one stock).

RESULTS

: The isolates presented typical T. cruzi morphology and usually grew well in routine culture media. Metacyclic trypomastigotes were found in cultures or experimentally infected Triatoma infestans. All isolates were pure T. cruzi cultures, presenting typical 330-bp products from kinetoplast DNA minicircles, and 250 or 200-bp amplicons from the mini-exon non-transcribed spacer. Their genetic type assignment was resolved by their isoenzyme profiles. The finding of TcI in one asymptomatic patient from Paraíba was confirmed by the sequencing assay. TcVI was found in two asymptomatic individuals from Bahia and Rio Grande do Sul. TcII was identified in six patients from Pernambuco, Bahia and Minas Gerais, who presented different clinical forms: cardiac (2), digestive with megaesophagus (1), and indeterminate (3).

CONCLUSIONS

: The main T. cruzi genotypes found in Brazilian chronic patients were identified in this work, including TcI, which is less frequent and usually causes asymptomatic disease, unlike that in other American countries. This study emphasizes the importance of T. cruzi genotyping for possible correlations between the parasite and patient' responses to therapeutic treatment or disease clinical manifestations.

摘要

引言

克氏锥虫是人类恰加斯病的病原体,主要流行于拉丁美洲。从在埃万德罗·恰加斯国家传染病研究所(巴西奥斯瓦尔多·克鲁兹基金会)接受随访的患者中通过血液培养分离出锥虫株,并采用不同方法进行研究。

方法

为进行物种和基因型鉴定,除了对一株克氏锥虫TcSC5D基因的多态性位点进行测序外,还通过寄生虫学技术、针对特定DNA序列的聚合酶链反应检测、同工酶谱分析对这些虫株进行分析。

结果

分离株呈现典型的克氏锥虫形态,通常在常规培养基中生长良好。在培养物或经实验感染的侵扰锥猎蝽中发现了后循环型锥鞭毛体。所有分离株均为纯克氏锥虫培养物,动质体DNA小环呈现典型的330bp产物,小外显子非转录间隔区呈现250或200bp扩增子。通过同工酶谱确定了它们的遗传类型。对来自帕拉伊巴州一名无症状患者的检测结果经测序分析证实为TcI型。在来自巴伊亚州和南里奥格兰德州的两名无症状个体中发现了TcVI型。在来自伯南布哥州、巴伊亚州和米纳斯吉拉斯州的6名患者中鉴定出TcII型,这些患者呈现不同临床类型:心脏型(2例)、伴有巨食管的消化型(1例)和隐匿型(3例)。

结论

本研究确定了巴西慢性病患者中主要的克氏锥虫基因型,包括TcI型,与其他美洲国家不同,该基因型较少见,通常导致无症状疾病。这项研究强调了克氏锥虫基因分型对于寄生虫与患者对治疗的反应或疾病临床表现之间可能存在的相关性的重要性。

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