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结合寄生虫学和基于分子的诊断工具可提高对恰加斯病患者单份外周血样本中克氏锥虫的检测。

Combined parasitological and molecular-based diagnostic tools improve the detection of Trypanosoma cruzi in single peripheral blood samples from patients with Chagas disease.

作者信息

Volpato Fabiana Caroline Zempulski, Sousa Giovane Rodrigo, D'Ávila Daniella Alchaar, Galvão Lúcia Maria da Cunha, Chiari Egler

机构信息

Programa de Pós-Graduação em Parasitologia, Departamento de Parasitologia, Instituto de Ciências Biológicas, Universidade Federal de Minas Gerais, Belo Horizonte, MG, Brasil.

Section on Immunobiology, Joslin Diabetes Center, Harvard Medical School, Boston, MA 02215, USA.

出版信息

Rev Soc Bras Med Trop. 2017 Jul-Aug;50(4):506-515. doi: 10.1590/0037-8682-0046-2017.

Abstract

INTRODUCTION

In order to detect Trypanosoma cruzi and determine the genetic profiles of the parasite during the chronic phase of Chagas disease (ChD), parasitological and molecular diagnostic methods were used to assess the blood of 91 patients without specific prior treatment.

METHODS

Blood samples were collected from 68 patients with cardiac ChD and 23 patients with an indeterminate form of ChD, followed by evaluation using blood culture and polymerase chain reaction. T . cruzi isolates were genotyped using three different genetic markers.

RESULTS

: Blood culture was positive in 54.9% of all patients, among which 60.3% had the cardiac form of ChD, and 39.1% the indeterminate form of ChD. There were no significant differences in blood culture positivity among patients with cardiac and indeterminate forms. Additionally, patient age and clinical forms did not influence blood culture results. Polymerase chain reaction (PCR) was positive in 98.9% of patients, although comparisons between blood culture and PCR results showed that the two techniques did not agree. Forty-two T . cruzi stocks were isolated, and TcII was detected in 95.2% of isolates. Additionally, one isolate corresponded to TcIII or TcIV, and another corresponded to TcV or TcVI.

CONCLUSIONS

Blood culture and PCR were both effective for identifying T. cruzi using a single blood sample, and their association did not improve parasite detection. However, we were not able to establish an association between the clinical form of ChD and the genetic profile of the parasite.

摘要

引言

为了检测克氏锥虫并确定恰加斯病(ChD)慢性期寄生虫的基因图谱,采用寄生虫学和分子诊断方法对91例未经特殊前期治疗的患者血液进行评估。

方法

采集68例心脏型ChD患者和23例不确定型ChD患者的血样,随后采用血培养和聚合酶链反应进行评估。使用三种不同的基因标记对克氏锥虫分离株进行基因分型。

结果

所有患者中血培养阳性率为54.9%,其中60.3%为心脏型ChD,39.1%为不确定型ChD。心脏型和不确定型患者的血培养阳性率无显著差异。此外,患者年龄和临床类型不影响血培养结果。聚合酶链反应(PCR)在98.9%的患者中呈阳性,尽管血培养和PCR结果比较显示这两种技术结果不一致。分离出42株克氏锥虫菌株,95.2%的分离株检测到TcII型。此外,一株分离株对应TcIII或TcIV型,另一株对应TcV或TcVI型。

结论

血培养和PCR均能通过单一血样有效鉴定克氏锥虫,二者联合使用并未提高寄生虫检测率。然而,我们未能确定ChD临床类型与寄生虫基因图谱之间的关联。

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