Hemi-nested PCR and RFLP methodologies for identifying blood meals of the Chagas disease vector, Triatoma infestans.

Dawn M. Roellig, Luis A. Gomez-Puerta, Daniel G. Mead, Jesus Pinto, Jenny Ancca-Juarez, Maritza Calderon, Caryn Bern, Robert H Gilman, Vitaliano A. Cama, Disease Workgroup in Arequipa Chagas Disease Workgroup in Arequipa

Research output: Contribution to journalArticle

Abstract

Trypanosoma cruzi, the etiologic agent of Chagas disease, is transmitted by hematophagous reduviid bugs within the subfamily Triatominae. These vectors take blood meals from a wide range of hosts, and their feeding behaviors have been used to investigate the ecology and epidemiology of T. cruzi. In this study we describe two PCR-based methodologies that amplify a fragment of the 16S mitochondrial rDNA, aimed to improve the identification of blood meal sources for Triatoma infestans: a.--Sequence analyses of two heminested PCRs that allow the identification of mammalian and avian species, and b.--restriction fragment length polymorphism (RFLP) analysis from the mammalian PCR to identify and differentiate multi-host blood meals. Findings from both methodologies indicate that host DNA could be detected and the host species identified in samples from laboratory reared and field collected triatomines. The implications of this study are two-fold. First, these methods can be used in areas where the fauna diversity and feeding behavior of the triatomines are unknown. Secondly, the RFLP method led to the identification of multi-host DNA from T. infestans gut contents, enhancing the information provided by this assay. These tools are important contributions for ecological and epidemiological studies of vector-borne diseases.

Original languageEnglish (US)
Article numbere74713
JournalPLoS One
Volume8
Issue number9
StatePublished - 2013
Externally publishedYes

Fingerprint

Triatoma
Triatoma infestans
Disease Vectors
disease vectors
Chagas disease
Chagas Disease
blood meal
Polymorphism
Triatominae
Restriction Fragment Length Polymorphisms
Meals
restriction fragment length polymorphism
Blood
Trypanosoma cruzi
Feeding Behavior
Polymerase Chain Reaction
feeding behavior
Epidemiology
Host Specificity
Ecology

ASJC Scopus subject areas

  • Agricultural and Biological Sciences(all)
  • Biochemistry, Genetics and Molecular Biology(all)
  • Medicine(all)

Cite this

Roellig, D. M., Gomez-Puerta, L. A., Mead, D. G., Pinto, J., Ancca-Juarez, J., Calderon, M., ... Chagas Disease Workgroup in Arequipa, D. W. I. A. (2013). Hemi-nested PCR and RFLP methodologies for identifying blood meals of the Chagas disease vector, Triatoma infestans. PLoS One, 8(9), [e74713].

Hemi-nested PCR and RFLP methodologies for identifying blood meals of the Chagas disease vector, Triatoma infestans. / Roellig, Dawn M.; Gomez-Puerta, Luis A.; Mead, Daniel G.; Pinto, Jesus; Ancca-Juarez, Jenny; Calderon, Maritza; Bern, Caryn; Gilman, Robert H; Cama, Vitaliano A.; Chagas Disease Workgroup in Arequipa, Disease Workgroup in Arequipa.

In: PLoS One, Vol. 8, No. 9, e74713, 2013.

Research output: Contribution to journalArticle

Roellig, DM, Gomez-Puerta, LA, Mead, DG, Pinto, J, Ancca-Juarez, J, Calderon, M, Bern, C, Gilman, RH, Cama, VA & Chagas Disease Workgroup in Arequipa, DWIA 2013, 'Hemi-nested PCR and RFLP methodologies for identifying blood meals of the Chagas disease vector, Triatoma infestans.', PLoS One, vol. 8, no. 9, e74713.
Roellig DM, Gomez-Puerta LA, Mead DG, Pinto J, Ancca-Juarez J, Calderon M et al. Hemi-nested PCR and RFLP methodologies for identifying blood meals of the Chagas disease vector, Triatoma infestans. PLoS One. 2013;8(9). e74713.
Roellig, Dawn M. ; Gomez-Puerta, Luis A. ; Mead, Daniel G. ; Pinto, Jesus ; Ancca-Juarez, Jenny ; Calderon, Maritza ; Bern, Caryn ; Gilman, Robert H ; Cama, Vitaliano A. ; Chagas Disease Workgroup in Arequipa, Disease Workgroup in Arequipa. / Hemi-nested PCR and RFLP methodologies for identifying blood meals of the Chagas disease vector, Triatoma infestans. In: PLoS One. 2013 ; Vol. 8, No. 9.
@article{f6d679bc1df24176886c8133f454eccb,
title = "Hemi-nested PCR and RFLP methodologies for identifying blood meals of the Chagas disease vector, Triatoma infestans.",
abstract = "Trypanosoma cruzi, the etiologic agent of Chagas disease, is transmitted by hematophagous reduviid bugs within the subfamily Triatominae. These vectors take blood meals from a wide range of hosts, and their feeding behaviors have been used to investigate the ecology and epidemiology of T. cruzi. In this study we describe two PCR-based methodologies that amplify a fragment of the 16S mitochondrial rDNA, aimed to improve the identification of blood meal sources for Triatoma infestans: a.--Sequence analyses of two heminested PCRs that allow the identification of mammalian and avian species, and b.--restriction fragment length polymorphism (RFLP) analysis from the mammalian PCR to identify and differentiate multi-host blood meals. Findings from both methodologies indicate that host DNA could be detected and the host species identified in samples from laboratory reared and field collected triatomines. The implications of this study are two-fold. First, these methods can be used in areas where the fauna diversity and feeding behavior of the triatomines are unknown. Secondly, the RFLP method led to the identification of multi-host DNA from T. infestans gut contents, enhancing the information provided by this assay. These tools are important contributions for ecological and epidemiological studies of vector-borne diseases.",
author = "Roellig, {Dawn M.} and Gomez-Puerta, {Luis A.} and Mead, {Daniel G.} and Jesus Pinto and Jenny Ancca-Juarez and Maritza Calderon and Caryn Bern and Gilman, {Robert H} and Cama, {Vitaliano A.} and {Chagas Disease Workgroup in Arequipa}, {Disease Workgroup in Arequipa}",
year = "2013",
language = "English (US)",
volume = "8",
journal = "PLoS One",
issn = "1932-6203",
publisher = "Public Library of Science",
number = "9",

}

TY - JOUR

T1 - Hemi-nested PCR and RFLP methodologies for identifying blood meals of the Chagas disease vector, Triatoma infestans.

AU - Roellig, Dawn M.

AU - Gomez-Puerta, Luis A.

AU - Mead, Daniel G.

AU - Pinto, Jesus

AU - Ancca-Juarez, Jenny

AU - Calderon, Maritza

AU - Bern, Caryn

AU - Gilman, Robert H

AU - Cama, Vitaliano A.

AU - Chagas Disease Workgroup in Arequipa, Disease Workgroup in Arequipa

PY - 2013

Y1 - 2013

N2 - Trypanosoma cruzi, the etiologic agent of Chagas disease, is transmitted by hematophagous reduviid bugs within the subfamily Triatominae. These vectors take blood meals from a wide range of hosts, and their feeding behaviors have been used to investigate the ecology and epidemiology of T. cruzi. In this study we describe two PCR-based methodologies that amplify a fragment of the 16S mitochondrial rDNA, aimed to improve the identification of blood meal sources for Triatoma infestans: a.--Sequence analyses of two heminested PCRs that allow the identification of mammalian and avian species, and b.--restriction fragment length polymorphism (RFLP) analysis from the mammalian PCR to identify and differentiate multi-host blood meals. Findings from both methodologies indicate that host DNA could be detected and the host species identified in samples from laboratory reared and field collected triatomines. The implications of this study are two-fold. First, these methods can be used in areas where the fauna diversity and feeding behavior of the triatomines are unknown. Secondly, the RFLP method led to the identification of multi-host DNA from T. infestans gut contents, enhancing the information provided by this assay. These tools are important contributions for ecological and epidemiological studies of vector-borne diseases.

AB - Trypanosoma cruzi, the etiologic agent of Chagas disease, is transmitted by hematophagous reduviid bugs within the subfamily Triatominae. These vectors take blood meals from a wide range of hosts, and their feeding behaviors have been used to investigate the ecology and epidemiology of T. cruzi. In this study we describe two PCR-based methodologies that amplify a fragment of the 16S mitochondrial rDNA, aimed to improve the identification of blood meal sources for Triatoma infestans: a.--Sequence analyses of two heminested PCRs that allow the identification of mammalian and avian species, and b.--restriction fragment length polymorphism (RFLP) analysis from the mammalian PCR to identify and differentiate multi-host blood meals. Findings from both methodologies indicate that host DNA could be detected and the host species identified in samples from laboratory reared and field collected triatomines. The implications of this study are two-fold. First, these methods can be used in areas where the fauna diversity and feeding behavior of the triatomines are unknown. Secondly, the RFLP method led to the identification of multi-host DNA from T. infestans gut contents, enhancing the information provided by this assay. These tools are important contributions for ecological and epidemiological studies of vector-borne diseases.

UR - http://www.scopus.com/inward/record.url?scp=84901815711&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=84901815711&partnerID=8YFLogxK

M3 - Article

C2 - 24040328

AN - SCOPUS:84901815711

VL - 8

JO - PLoS One

JF - PLoS One

SN - 1932-6203

IS - 9

M1 - e74713

ER -