Grupo Biología y Control de Enfermedades Infecciosas (BCEI), Sede de Investigación Universitaria, Universidad de Antioquia, Medellín, Colombia.
PLoS Negl Trop Dis. 2012;6(2):e1530. doi: 10.1371/journal.pntd.0001530. Epub 2012 Feb 28.
Methods to determine blood-meal sources of hematophagous Triatominae bugs (Chagas disease vectors) are serological or based on PCR employing species-specific primers or heteroduplex analysis, but these are expensive, inaccurate, or problematic when the insect has fed on more than one species. To solve those problems, we developed a technique based on HRM analysis of the mitochondrial gene cytochrome B (Cyt b). This technique recognized 14 species involved in several ecoepidemiological cycles of the transmission of Trypanosoma cruzi and it was suitable with DNA extracted from intestinal content and feces 30 days after feeding, revealing a resolution power that can display mixed feedings. Field samples were analyzed showing blood meal sources corresponding to domestic, peridomiciliary and sylvatic cycles. The technique only requires a single pair of primers that amplify the Cyt b gene in vertebrates and no other standardization, making it quick, easy, relatively inexpensive, and highly accurate.
方法来确定血食源吸血三锥虫（恰加斯病媒介）是血清学或基于 PCR 采用种特异性引物或异源双链分析，但这些都是昂贵的，不准确的，或有问题时，昆虫已经喂了不止一个物种。为了解决这些问题，我们开发了一种基于 HRM 分析线粒体基因细胞色素 B（Cyt b）的技术。该技术识别了 14 种参与克氏锥虫传播的几个生态流行病学循环的物种，并且适用于从肠道内容物和喂食后 30 天的粪便中提取的 DNA，显示出能够显示混合喂养的分辨率能力。对现场样本进行了分析，显示出与家庭、围屋和森林循环相对应的血液摄入源。该技术只需要一对引物，在脊椎动物中扩增 Cyt b 基因，不需要其他标准化，因此快速、简单、相对便宜、高度准确。
