high-resolution melting (hrm) of the cytochrome b gene a powerful approach to identify blood-meal sources in chagas disease vectors高分辨率融化(人力资源管理)的细胞色素b基因的一个强大的方法来识别血源恰加斯病向量.pdfVIP

High-Resolution Melting (HRM) of the Cytochrome B Gene: A Powerful Approach to Identify Blood-Meal Sources in Chagas Disease Vectors ˜ ´ ´ ´ ´ Victor H. Pena, Geysson J. Fernandez, Andres M. Gomez-Palacio, Ana M. Mejıa-Jaramillo, Omar Cantillo, ´ Omar Triana-Chavez* ´ ´ ´ Grupo Biologıa y Control de Enfermedades Infecciosas (BCEI), Sede de Investigacion Universitaria, Universidad de Antioquia, Medellın, Colombia Abstract 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. ˜ ´ ´ ´ Citation: Pena VH, Fernandez GJ, Gomez-Palacio AM, Mejıa-Jaramillo AM, Cantillo O, et al. (2012) High-Resol