Diseño y evaluación de tres oligonucleótidos para la detección de Leishmania por PCR

Abstract

Leishmaniasis affects public health in 88 countries of the world and represents an important obstacle for their socioeconomic development. The methods used to diagnose this disease take time and are frequently traumatic to the patient. Materials and methods Genomic DNA from different Leishmania species were extracted. The carboxi-end terminal of a partial sequence of the H2B histone of Leishmania (V.) peruviana was used to design three oligonucleotides (LEISH 1, LEISH 2, LEISH3) to design a PCR technique as an alternative method to diagnose this disease rapidly. Results: A PCR was performed with these primers, which amplified succesfully 123bp (LEISH 1/ LEISH 2) and 139bp (LEISH1/ LEISH3) of the partial sequence studied. The sensitivity of direct PCR was appropiate even when 1fg of purified DNA of L. (V.) peruviana and 2 parasites was used. The specificity was 100% in all Leishmania species analyzed and did not recognized neither Trypanosoma nor human DNA. Conclusions: The oligonucleotides designed could be used in the detection of parasite from different kind of samples like fresh blood with anticoagulant, biopsy and dermal scrapping. PCR has become an alternative method to diagnose leishmaniasis because of its speed, specificity and sensitivity.