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Sources of blood meals of sylvatic Triatoma guasayana near Zurima, Bolivia, assayed with qPCR and 12S cloning.

Lucero DE, Ribera W, Pizarro JC, Plaza C, Gordon LW, Peña R, Morrissey LA, Rizzo DM, Stevens L - PLoS Negl Trop Dis (2014)

Bottom Line: 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.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.

View Article: PubMed Central - PubMed

Affiliation: Department of Biology, University of Vermont, Burlington, Vermont, United States of America; Vector-borne Diseases Section, Tennessee Department of Health, Nashville, Tennessee, United States of America.

ABSTRACT

Background: In 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 findings: We 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/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. 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.

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Related in: MedlinePlus

Satellite imagery (∼1 m resolution, ArcGIS, Ver. 10.1, ESRI Inc., Redlands, California, USA [26]) centered on the riverbed of Rio Chico near the town of Zurima (left).Digitized town boundary (black polygon), riverbed (blue lines), six isolated houses (small black dots) and numbered locations of the positive (red fill) and negative (white fill) trap sites.
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pntd-0003365-g001: Satellite imagery (∼1 m resolution, ArcGIS, Ver. 10.1, ESRI Inc., Redlands, California, USA [26]) centered on the riverbed of Rio Chico near the town of Zurima (left).Digitized town boundary (black polygon), riverbed (blue lines), six isolated houses (small black dots) and numbered locations of the positive (red fill) and negative (white fill) trap sites.

Mentions: Located in the Andean highlands of the department of Chuquisaca, Bolivia, the rural landscape of the study area includes xeric valleys [22], with thorny shrubs and cacti adapted to low, seasonal rainfall that defines the dry (8.4 cm average rainfall of May to October) and wet seasons (35.8 cm average rainfall of December to March) [23]. The sampling locations (Mean Center calculated at 65o 08′ 2.31″ W, 18o 46′ 44.26″ with ArcGIS, Ver. 10.1, ESRI Inc., Redlands, California, USA) were within 200 m of Rio Chico; six sampling locations were evenly divided between the west and east banks. Elevation of these locations range between 1740 m and 1780 m above mean sea level (AMSL, Figure 1).


Sources of blood meals of sylvatic Triatoma guasayana near Zurima, Bolivia, assayed with qPCR and 12S cloning.

Lucero DE, Ribera W, Pizarro JC, Plaza C, Gordon LW, Peña R, Morrissey LA, Rizzo DM, Stevens L - PLoS Negl Trop Dis (2014)

Satellite imagery (∼1 m resolution, ArcGIS, Ver. 10.1, ESRI Inc., Redlands, California, USA [26]) centered on the riverbed of Rio Chico near the town of Zurima (left).Digitized town boundary (black polygon), riverbed (blue lines), six isolated houses (small black dots) and numbered locations of the positive (red fill) and negative (white fill) trap sites.
© Copyright Policy
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC4256209&req=5

pntd-0003365-g001: Satellite imagery (∼1 m resolution, ArcGIS, Ver. 10.1, ESRI Inc., Redlands, California, USA [26]) centered on the riverbed of Rio Chico near the town of Zurima (left).Digitized town boundary (black polygon), riverbed (blue lines), six isolated houses (small black dots) and numbered locations of the positive (red fill) and negative (white fill) trap sites.
Mentions: Located in the Andean highlands of the department of Chuquisaca, Bolivia, the rural landscape of the study area includes xeric valleys [22], with thorny shrubs and cacti adapted to low, seasonal rainfall that defines the dry (8.4 cm average rainfall of May to October) and wet seasons (35.8 cm average rainfall of December to March) [23]. The sampling locations (Mean Center calculated at 65o 08′ 2.31″ W, 18o 46′ 44.26″ with ArcGIS, Ver. 10.1, ESRI Inc., Redlands, California, USA) were within 200 m of Rio Chico; six sampling locations were evenly divided between the west and east banks. Elevation of these locations range between 1740 m and 1780 m above mean sea level (AMSL, Figure 1).

Bottom Line: 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.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.

View Article: PubMed Central - PubMed

Affiliation: Department of Biology, University of Vermont, Burlington, Vermont, United States of America; Vector-borne Diseases Section, Tennessee Department of Health, Nashville, Tennessee, United States of America.

ABSTRACT

Background: In 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 findings: We 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/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. 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.

Show MeSH
Related in: MedlinePlus