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用于检测滤纸收集的骚扰锥蝽干粪便斑点中克氏锥虫的快速DNA分离和扩增方法的适用性。

Suitability of a rapid DNA isolation and amplification for detection of Trypanosoma cruzi in Triatoma infestans dry fecal spots collected on filter paper.

作者信息

Braz L M A, Raiz-Júnior R, Alárcon R S, Gakiya E, Amato-Neto V, Okay T S

机构信息

LIM 36, Laboratório de Investigação Médica, Pediatria Clínica, Departamento de Pediatria, Faculdade de Medicina da Universidade de São Paulo, Brasil.

出版信息

Parasite. 2008 Dec;15(4):595-8. doi: 10.1051/parasite/2008154595.

Abstract

A rapid DNA extraction was used for T. cruzi detection in triatomines dry fecal spots collected on filter paper and analyzed by PCR. Fifty T. infestans were fed on experimentally infected Balb/C mice with high T. cruzi parasitemia and divided into five groups of ten triatomines, and 100 triatomines were infected with lower parasitemia and divided into five groups of 20 triatomines. One dry fecal spot was analyzed per group on days 1, 2, 3, 4 and 5 post feeding. Amplification targeted T. cruzi TCZ sequence and resulted positive from day 4 after bugs feeding in the two models (high and lower parasitemial. The rapid DNA isolation and PCR proposed are suitable for detection of T. cruzi DNA in filter paper and should be considered in field research.

摘要

采用快速DNA提取法,对滤纸上收集的锥蝽干粪便斑点进行克氏锥虫检测,并通过聚合酶链反应(PCR)进行分析。五十只侵扰锥蝽以患有高克氏锥虫血症的经实验感染的Balb/C小鼠为食,并分为五组,每组十只锥蝽;一百只锥蝽感染了较低的虫血症,并分为五组,每组二十只锥蝽。在喂食后的第1、2、3、4和5天,每组分析一个干粪便斑点。扩增靶向克氏锥虫TCZ序列,在两种模型(高虫血症和低虫血症)中,锥蝽喂食后第4天结果呈阳性。所提出的快速DNA分离和PCR方法适用于检测滤纸上的克氏锥虫DNA,应在现场研究中予以考虑。

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