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Clinical and Diagnostic Laboratory Immunology, September 2003, p. 775-779, Vol. 10, No. 5
1071-412X/03/$08.00+0     DOI: 10.1128/CDLI.10.5.775-779.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.

Detection of Trypanosoma cruzi and Trypanosoma rangeli Infection by Duplex PCR Assay Based on Telomeric Sequences

Miguel Angel Chiurillo,1,2 Gladys Crisante,3 Agustina Rojas,3 Andreina Peralta,4 Manuel Dias,4 Palmira Guevara,4 Néstor Añez,3 and José Luis Ramírez2,4*

Decanato de Medicina, Universidad Centroccidental "Lisandro Alvarado," Barquisimeto 3001,1 Instituto de Estudios Avanzados, Ministerio de Ciencia y Tecnología, Caracas,2 Investigaciones Parasitológicas "José Francisco Torrealba," Universidad de Los Andes, Mérida,3 Instituto de Biología Experimental, Universidad Central de Venezuela, 1041-A Caracas, Venezuela4

Received 30 January 2003/ Returned for modification 31 March 2003/ Accepted 10 June 2003

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.

* Corresponding author. Mailing address: IBE-UCV Calle Suapure, Colinas de Bello Monte, P.O. Box Apdo 47525, Caracas 1041-A, Venezuela. Phone: 58 212 9621605. Fax: 58 212 9621120. E-mail address: jramirez{at}reacciun.ve.

Clinical and Diagnostic Laboratory Immunology, September 2003, p. 775-779, Vol. 10, No. 5
1071-412X/03/$08.00+0     DOI: 10.1128/CDLI.10.5.775-779.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.





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