Protein mass spectrometry extends temporal blood meal detection over polymerase chain reaction in mouse-fed Chagas disease vectors

Abstract

BACKGROUND Chagas disease is highly prevalent in Latin America, and vector control is the most effective control strategy to date. We have previously shown that liquid chromatography tandem mass spectrometry (LC-MS/MS) is a valuable tool for identifying triatomine vector blood meals.OBJECTIVES The purpose of this study was to determine blood meal detection ability as a function of method [polymerase chain reaction (PCR) vs. LC-MS/MS], time since feeding, and the effect of molting in mouse-fed triatomine insect vectors targeting hemoglobin and albumin proteins with LC-MS/MS and short interspersed nuclear elements (SINE)-based PCR.METHODS We experimentally fed Triatoma protracta on mice and used LC-MS/MS to detect hemoglobin and albumin peptides over time post-feeding and post-molting (≤ 12 weeks). We compared LC-MS/MS results with those of a standard PCR method based on SINEs.FINDINGS Hemoglobin-based LC-MS/MS detected blood meals most robustly at all time points post-feeding. Post-molting, no blood meals were detected with PCR, whereas LC-MS/MS detected mouse hemoglobin and albumin up to 12 weeks.MAIN CONCLUSIONS In our study, the hemoglobin signature in the insect abdomen lasted longer than that of albumin and DNA. LC-MS/MS using hemoglobin shows promise for identifying triatomine blood meals over long temporal scales and even post-molting. Clarifying the frequency of blood-feeding on different hosts can foster our understanding of vector behavior and may help devise sounder disease-control strategies, including Ecohealth (community based ecosystem management) approaches.

Rapid detection of human blood in triatomines (kissing bugs) utilizing a lateral flow immunochromatographic assay - A pilot study

“…All procedures using mice were approved by the Southwestern Biological Institute, Tucson, Arizona, USA Animal Care and Use Committee following international standards. 7 Human venous blood was provided from a volunteer after informed consent.…”

“…This triatomine colony originates from a 2009 collection of wild-caught T. rubida from Tucson, Arizona, USA. The T. rubida colony is maintained as described by Keller et al 7 Fifteen laboratory-raised fifth instar T. rubida were fed mouse blood for 30-60 min or until engorgement, at which time the mouse was removed. Five laboratory-raised fifth instar T. rubida having only fed previously on mouse blood (approximately six weeks prior) were fed human blood through an artificial membrane feeding apparatus.…”

“… 2 Several techniques have been developed using DNA- and proteomics-based methodologies in an attempt to determine the blood meal sources of triatomines. 3 , 4 , 5 , 6 , 7 These procedures are sensitive and specific for identifying a blood meal source, but are not readily adaptable to field use. They require access to a laboratory with sophisticated equipment and trained personnel, are labor intensive, and are vulnerable to contamination and degradation of the blood meal.…”

“…They require access to a laboratory with sophisticated equipment and trained personnel, are labor intensive, and are vulnerable to contamination and degradation of the blood meal. 3 , 4 , 5 , 6 , 7 The Rapid Stain Identification of Human Blood (RSID™ Blood) is a lateral flow, immunochromatographic assay detecting as little as 1 µL of human blood in forensic samples within 10 min following 1-2 h of specimen preparation. We conducted a preliminary study using this assay on triatomines and their excreta fed mouse or human blood and also numerous field specimens.…”

Implementation science: Epidemiology and feeding profiles of the Chagas vector Triatoma dimidiata prior to Ecohealth intervention for three locations in Central America

“…As mentioned before, the lack of blood source detection by PCR can indicate either a recent blood meal from taxa not included in the survey or no recent blood meal [4,20,41]. Strong support for no recent blood meal is provided by recent studies based on mass spectrometry [54,55] including domestic vectors from El Salvador that show DNA based methods have a short window for blood meal detection [54], it would be interesting to examine T. dimidiata where no blood sources were detected by PCR from these three localities to strengthen the Ecohealth strategies proposed by this study.…”