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通过实时定量聚合酶链反应鉴定克氏锥虫谱系并确定墨西哥锥蝽肠道样本中的血餐来源

Identification by Q-PCR of Trypanosoma cruzi lineage and determination of blood meal sources in triatomine gut samples in México.

作者信息

Ibáñez-Cervantes Gabriela, Martínez-Ibarra Alejandro, Nogueda-Torres Benjamín, López-Orduña Eduardo, Alonso Ana L, Perea Cynthia, Maldonado Teresa, Hernández José Manuel, León-Avila Gloria

机构信息

Departamento de Zoología, Escuela Nacional de Ciencias Biológicas, Instituto Politécnico Nacional, Prol. Carpio y Plan de Ayala s/n, Col. Santo Tomás, CP 11340, México, DF, México.

出版信息

Parasitol Int. 2013 Feb;62(1):36-43. doi: 10.1016/j.parint.2012.09.003. Epub 2012 Sep 17.

Abstract

Triatomine vectors were collected on human dwellings in Michoacán México. Blood meal sources were identified by real time polymerase chain reaction (Q-PCR) using DNA extracted from triatomine guts. The assay was performed with one only specific primer set to amplify a fragment of the mitochondrial 12S ribosomal gene from vertebrate species. Also Trypanosoma cruzi parasites were detected in triatomine gut samples by microscopy and the positive infection was tested in mice. In addition T. cruzi discrete taxonomic units (DTUs) were identified by Q-PCR with two sets of primers that amplify the mini-circle region (miniexon) and 18S ribosomal mitochondrial gene. The sequences obtained from 18S ribosomal gene amplifications confirmed the presence of T. cruzi I and II lineages, and provide evidence of the presence of lineage TcIII and TcIV.

摘要

在墨西哥米却肯州的人类住所采集了锥蝽媒介。通过实时聚合酶链反应(Q-PCR),利用从锥蝽肠道中提取的DNA来鉴定血餐来源。该检测使用一组特异性引物进行,以扩增脊椎动物物种线粒体12S核糖体基因的片段。此外,通过显微镜在锥蝽肠道样本中检测克氏锥虫寄生虫,并在小鼠身上测试阳性感染情况。另外,使用两组引物通过Q-PCR鉴定克氏锥虫离散分类单元(DTUs),这两组引物可扩增小环区域(小外显子)和18S核糖体线粒体基因。从18S核糖体基因扩增获得的序列证实了克氏锥虫I和II谱系的存在,并提供了TcIII和TcIV谱系存在的证据。

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