Abstract
BackgroundIn this study we compared the utility of two molecular biology techniques, cloning of the mitochondrial 12S ribosomal RNA gene and hydrolysis probe-based qPCR, to identify blood meal sources of sylvatic Chagas disease insect vectors collected with live-bait mouse traps (also known as Noireau traps). Fourteen T. guasayana were collected from six georeferenced trap locations in the Andean highlands of the department of Chuquisaca, Bolivia.Methodology/Principal FindingsWe detected four blood meals sources with the cloning assay: seven samples were positive for human (Homo sapiens), five for chicken (Gallus gallus) and unicolored blackbird (Agelasticus cyanopus), and one for opossum (Monodelphis domestica). Using the qPCR assay we detected chicken (13 vectors), and human (14 vectors) blood meals as well as an additional blood meal source, Canis sp. (4 vectors).Conclusions/SignificanceWe show that cloning of 12S PCR products, which avoids bias associated with developing primers based on a priori knowledge, detected blood meal sources not previously considered and that species-specific qPCR is more sensitive. All samples identified as positive for a specific blood meal source by the cloning assay were also positive by qPCR. However, not all samples positive by qPCR were positive by cloning. We show the power of combining the cloning assay with the highly sensitive hydrolysis probe-based qPCR assay provides a more complete picture of blood meal sources for insect disease vectors.
Highlights
Blood-feeding insects in the subfamily Triatominae are vectors of Trypanosoma cruzi, the parasite that causes Chagas disease
Conclusions/Significance: We show that cloning of 12S PCR products, which avoids bias associated with developing primers based on a priori knowledge, detected blood meal sources not previously considered and that species-specific qPCR is more sensitive
We show the power of combining the cloning assay with the highly sensitive hydrolysis probe-based qPCR assay provides a more complete picture of blood meal sources for insect disease vectors
Summary
Blood-feeding insects in the subfamily Triatominae are vectors of Trypanosoma cruzi, the parasite that causes Chagas disease. 140 species of Triatominae range across the Americas [1] and vary in their role in transmitting human Chagas disease in part because of their preference for sylvatic (wild), peridomestic (immediate surroundings of a house) or domestic (within house) ecotopes. Because sylvatic vectors have the potential to colonize houses or enter houses to feed and leave, collecting sylvatic vectors and analyzing their blood meal profiles can provide insight into their movement among domestic, peridomestic and sylvatic ecotopes and their potential role in disease transmission [2]. Vectors that feed on human blood are important in disease transmission but because some mammals are more likely to transmit the parasite to the vectors, insight into the spectrum of blood meal sources is epidemiologically important.
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