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Geographical clustering of Trypanosoma cruzi I groups from Colombia revealed by low-stringency single specific primer-PCR of the intergenic regions of spliced-leader genes

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Abstract

A low-stringency single-primer polymerase chain reaction (LSSP-PCR) typing procedure targeted to the intergenic regions of spliced-leader genes (SL) was designed to profile Trypanosoma cruzi I stocks from endemic regions of Colombia. Comparison between SL-LSSP-PCR profiles of parasite DNA from vector faeces and cultures isolated from those faeces showed more conservative signatures than profiles using LSSP-PCR targeted to the minicircle variable regions (kDNA). This was also observed by analysing 15 parasite clones from one stock as well as serial samples of a same stock after in vitro culturing or inoculation into mice. Thus, SL-LSSP-PCR appears more appropriate than kDNA-LSSP-PCR for reliable typing of major T. cruzi I groups from in vitro cultured stocks and triatomine faeces. SL-LSSP-PCR grouped 46 of 47 T. cruzi I Colombian stocks according to their geographical procedences in four clusters: Cluster Cas from Casanare Department, Cluster Mg from Northern Magdalena department, Cluster Mom from Momposina Depression in Southern Magdalena and finally Cluster NW from northwestern Colombia, including Sucre, Chocó, Córdoba and Antioquia departments. Sequence analysis identified punctual mutations among amplicons from each cluster. Within Cluster Mg, sequence polymorphism allowed association with different sylvatic vector species. Novel SL sequences and LSSP-PCR profiles are reported from T. cruzi I infecting Eratyrus cuspidatus, Panstrongylus geniculatus and Rhodnius pallescens vectors.

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Acknowledgements

This work was financed by Colciencias cod 1115-04-14387; Proyecto de Sostenibilidad, University of Antioquia 2007–2008 to OTC; Proyecto 2020, Banco de la República of Colombia to AMJ and OTC, and partially by PIP 5469 CONICET and PICT 33955 to AGS. All experimental procedures comply with the criteria of the Ethical Committee of the University of Antioquia and INGEBI-CONICET, in agreement with the current laws of the corresponding countries.

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  1. Grupo de Chagas, Universidad de Antioquia, AA 1226, Medellin, Colombia

    Ana María Mejía-Jaramillo, Sair Arboleda-Sánchez, Ingrid Bibiana Rodríguez, Alexander Salazar & Omar Triana-Chávez

  2. Centro de Investigaciones Biológicas, (CSIC), Madrid, Spain

    Jesús del Mazo

  3. Laboratorio de Biología Molecular de la Enfermedad de Chagas, INGEBI-CONICET, Buenos Aires, Argentina

    Carolina Cura & Alejandro Gabriel Schijman

  4. Vuelta de Obligado 2490, Buenos Aires, 1428, Argentina

    Alejandro Gabriel Schijman

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  1. Ana María Mejía-Jaramillo
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Mejía-Jaramillo, A.M., Arboleda-Sánchez, S., Rodríguez, I.B. et al. Geographical clustering of Trypanosoma cruzi I groups from Colombia revealed by low-stringency single specific primer-PCR of the intergenic regions of spliced-leader genes. Parasitol Res 104, 399–410 (2009). https://doi.org/10.1007/s00436-008-1212-0

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