title: An alternative approach to detect Trypanosoma in Glossina (Diptera, Glossinidae) without dissection.
abstract: BACKGROUND
  Determining if a tsetse fly is infected by trypanosomes and thus potentially able to transmit trypanosome-related human and animal diseases is an extremely laborious and time-consuming task to perform, especially under field conditions. In this study we tested a possible alternative approach that uses the entire insect vector for DNA extraction and PCR analysis to detect and identify Trypanosoma spp. in field collected tsetse flies.
  
  
  METHODS
  DNA extraction was performed using a method originally developed for tick DNA extraction followed by PCR detection and identification of Trypanosoma spp.
  
  
  RESULTS
  Two out of 62 flies captured in Equatorial Guinea carried DNA of T. brucei s.l. and Trypanosoma vivax. T. congolense forest, T. congolense savannah and T. congolense Kilifi were not detected.
  
  
  CONCLUSIONS
  The approach we employed allowed the molecular detection and species identification of trypanosomes using the whole vector body for DNA extraction. Although the approach does not give direct information on tsetse infectivity, it provides valuable information about trypanosome species circulating in a tsetse fly vector population. The method allows an effective processing of a large number of field captured tsetse in a central laboratory.
authors.affiliation: Unidade de Clínica das Doenças Tropicais and Centro de Malária e Outras Doenças Tropicais - LA, Instituto de Higiene e Medicina Tropical, Universidade Nova de Lisboa. Rua da Junqueira, 96, 1349-00 Lisboa, Portugal.
authors.forename: Filipa
authors.initials: F
authors.lastname: Ferreira
authors.forename: Jorge
authors.initials: J
authors.lastname: Cano
authors.forename: Andreia
authors.initials: A
authors.lastname: Furtado
authors.forename: Nicolas
authors.initials: N
authors.lastname: Ndong-Mabale
authors.forename: Pedro
authors.initials: P
authors.lastname: Ndong-Asumu
authors.forename: Agustin
authors.initials: A
authors.lastname: Benito
authors.forename: João
authors.initials: J
authors.lastname: Pinto
authors.forename: Maria Odete
authors.initials: MO
authors.lastname: Afonso
authors.forename: Jorge
authors.initials: J
authors.lastname: Seixas
authors.forename: Jorge
authors.initials: J
authors.lastname: Atouguia
authors.forename: Sónia
authors.initials: S
authors.lastname: Centeno-Lima
mesh_terms: D000818:Animals; D017931:DNA Primers / Q000235:genetics; D016703:Equatorial Guinea; D007303:Insect Vectors; D016133:Polymerase Chain Reaction / Q000379:methods*; D011159:Population Surveillance; D014345:Trypanosoma / Q000145:classification / Q000235:genetics* / Q000302:isolation & purification; D014370:Tsetse Flies / Q000469:parasitology*
chemical_list: D017931:DNA Primers