Centers for Disease Control and Prevention, Division of Parasitic Diseases and Malaria, Atlanta, Georgia, United States of America.
PLoS One. 2013 Sep 11;8(9):e74713. doi: 10.1371/journal.pone.0074713. eCollection 2013.
Trypanosoma cruzi, the etiologic agent of Chagas disease, is transmitted by hematophagous reduviid bugs within the subfamily Triatominae. These vectors take blood meals from a wide range of hosts, and their feeding behaviors have been used to investigate the ecology and epidemiology of T. cruzi. In this study we describe two PCR-based methodologies that amplify a fragment of the 16S mitochondrial rDNA, aimed to improve the identification of blood meal sources for Triatoma infestans: a.--Sequence analyses of two heminested PCRs that allow the identification of mammalian and avian species, and b.--restriction fragment length polymorphism (RFLP) analysis from the mammalian PCR to identify and differentiate multi-host blood meals. Findings from both methodologies indicate that host DNA could be detected and the host species identified in samples from laboratory reared and field collected triatomines. The implications of this study are two-fold. First, these methods can be used in areas where the fauna diversity and feeding behavior of the triatomines are unknown. Secondly, the RFLP method led to the identification of multi-host DNA from T. infestans gut contents, enhancing the information provided by this assay. These tools are important contributions for ecological and epidemiological studies of vector-borne diseases.
克氏锥虫，恰加斯病的病原体，通过在三锥虫亚科内的吸血蝽传播。这些媒介从广泛的宿主身上吸血，它们的进食行为被用来研究克氏锥虫的生态学和流行病学。在这项研究中，我们描述了两种基于 PCR 的方法，扩增 16S 线粒体 rDNA 的一个片段，旨在提高对侵染锥虫的血食来源的鉴定：a.——两个半巢式 PCR 的序列分析，允许鉴定哺乳动物和鸟类物种，和 b.——来自哺乳动物 PCR 的限制性片段长度多态性（RFLP）分析，以鉴定和区分多宿主血食。这两种方法的结果都表明，在实验室饲养和野外采集的锥虫样本中，可以检测到宿主 DNA 并鉴定出宿主物种。这项研究的意义有两方面。首先，这些方法可以用于那些三锥虫动物群多样性和进食行为未知的地区。其次，RFLP 方法从侵染锥虫的肠道内容物中鉴定出了多宿主 DNA，增强了该检测方法提供的信息。这些工具是对虫媒疾病的生态学和流行病学研究的重要贡献。
