Detection of Trypanosoma cruzi and Trypanosoma rangeli infection by duplex PCR assay based on telomeric sequences.
We used the species specificity and repetitious nature of subtelomeric kinetoplastida sequences to generate a duplex PCR assay for the simultaneous detection of Trypanosoma cruzi and Trypanosoma rangeli in experimentally and naturally infected triatomine (Reduviid) bugs and in infected human subjects. The assay was species specific and was capable of detecting 1/20th of T. cruzi and 1/4th of T. rangeli cell equivalents without complementary hybridization. In addition, the PCR-based assay was robust enough for direct application to difficult biological samples such as Reduviid feces or guts and was capable of recognizing all T. cruzi and T. rangeli strains and lineages. Because the assay primers amplify entirely different target sequences, no reaction interference was observed, facilitating future adaptation of this assay to an automated format.
Chiurillo
Miguel Angel
MA
Decanato de Medicina, Universidad Centroccidental Lisandro Alvarado, Barquisimeto 3001, Venezuela.
Crisante
Gladys
G
Rojas
Agustina
A
Peralta
Andreina
A
Dias
Manuel
M
Guevara
Palmira
P
Añez
Néstor
N
Ramírez
José Luis
JL
D000818:Animals; D017931:DNA Primers; D006801:Humans; D016133:Polymerase Chain Reaction / Q000379:methods*; D020064:Reduviidae / Q000469:parasitology; D012680:Sensitivity and Specificity; D013045:Species Specificity; D014345:Trypanosoma / Q000276:immunology* / Q000302:isolation & purification; D014352:Trypanosomiasis / Q000175:diagnosis* / Q000276:immunology
D017931:DNA Primers