Abstract
BackgroundIn a SINE-based PCR assay, a primer set specific for guinea pig genome short interspersed elements DNA was used to test the utility of genomic markers for identifying the source of vertebrate blood meals of Triatoma infestans.MethodsThe investigation consisted of two assays. In Assay 1, thirty-six insects, collected from the Province of Zudáñez in Chuquisaca, Bolivia were frozen 1–40 hours after feeding, under controlled conditions, on guinea pigs. The species of the vertebrate host was confirmed from dissection of the posterior part of the abdomen of each insect followed by DNA extraction and PCR amplification. Assay 2 investigated whether the technique worked under field conditions. We analyzed the bloodmeal of 34 insects collected from households and peri-domestic structures from communities where wild and captive guinea pigs occur. After collection, the insects were maintained at room temperature for 2 months without feeding and then analyzed.ResultsIn Assay 1, each of the 36 insects allowed to feed on guinea pig blood tested positive for guinea pig DNA. The guinea pig DNA was reliably identified in as little as 1 hour and up to 40 hours after feeding. For Assay 2, 8 out of the 34 samples (23%) showed positive results with guinea pig specific primers.ConclusionThe results in assay 1 demonstrated that DNA from the vertebrate host can be amplified 1–40 hours post feeding from the abdomen of the blood-feeding Chagas disease vector Triatoma infestans. The results in assay 2 confirmed that the procedure works on insects collected from households and peri-domestic structures and that the source of a blood meal can be determined at least 2 months post feeding.
Highlights
In a SINE-based Polymerase chain reaction (PCR) assay, a primer set specific for guinea pig genome short interspersed elements DNA was used to test the utility of genomic markers for identifying the source of vertebrate blood meals of Triatoma infestans
Comparison of the sequence of the PCR products to those in GenBank confirmed that the amplified product was guinea pig DNA
Some of the negative samples were spiked with 0.3 ng guinea pig DNA and subsequent PCR with guinea pig specific primers showed amplification in all cases
Summary
In a SINE-based PCR assay, a primer set specific for guinea pig genome short interspersed elements DNA was used to test the utility of genomic markers for identifying the source of vertebrate blood meals of Triatoma infestans. The recommended methods of disease control include screening blood donations, eliminating domestic vector populations by plastering adobe walls and using tile roofs to reduce vector habitat in homes, and spraying infested houses with residual insecticides [4] These methods, effective through much of Central and South America, have not been successful in Bolivia where prevalence of human infection and house infestation by the main vector in this area, Triatoma infestans, remains high. Identification of the source of the blood meal in the digestive tract of the hematophagous triatomines that transmit Chagas disease provides data on host-feeding patterns and preferences in nature These data provide information about epidemiologically significant disease reservoirs for the most serious human parasitic disease of the Americas in terms of social and economic impact [10]
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