Specific primers design based on the superoxide dismutase b gene for Trypanosoma cruzi as a screening tool: Validation method using strains from Colombia classified according to their discrete typing unit.
Olmo, Francisco;
Escobedo-Orteg, Javier;
Palma, Patricia;
Sánchez-Moreno, Manuel;
Mejía-Jaramillo, Ana;
Triana, Omar;
Marín, Clotilde;
(2014)
Specific primers design based on the superoxide dismutase b gene for Trypanosoma cruzi as a screening tool: Validation method using strains from Colombia classified according to their discrete typing unit.
Asian Pac J Trop Med, 7 (11).
pp. 854-859.
ISSN 2352-4146
DOI: https://doi.org/10.1016/S1995-7645(14)60149-8
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OBJECTIVE: To classify 21 new isolates of Trypanosoma cruzi (T. cruzi) according to the Discrete Typing Unit (DTU) which they belong to, as well as tune up a new pair of primers designed to detect the parasite in biological samples. METHODS: Strains were isolated, DNA extracted, and classified by using three Polymerase Chain Reactions (PCR). Subsequently this DNA was used along with other isolates of various biological samples, for a new PCR using primers designed. Finally, the amplified fragments were sequenced. RESULTS: It was observed the predominance of DTU I in Colombia, as well as the specificity of our primers for detection of T. cruzi, while no band was obtained when other species were used. CONCLUSIONS: This work reveals the genetic variability of 21 new isolates of T. cruzi in Colombia.Our primers confirmed their specificity for detecting the presence of T. cruzi.