RAPD method useful for distinguishing Leishmania species: design of specific primers for L. braziliensis

Abstract

The technique of Random Amplification Polymorphic DNA allows fragments of the genome to be amplified by means of polymerase chain reaction (PCR) without previous knowledge of their sequences. The protozoa of the genus Leishmania present great genetic variability, making it difficult to characterize the different species. A method is developed with a single 10-mers long primer, which allows the species L. braziliensis, L. mexicana, L. infantum, L. tropica, L. chagasi, L. amazonensis and L. major to be differentiated. These products amplified by RAPD have also facilitated the design of some primers that amplify L. braziliensis DNA exclusively.

Lutzomyia longipalpis and the eco-epidemiology of American visceral leishmaniasis, with particular reference to Brazil: a review

“…More recently Lainson and Rangel (2003) and Lainson and Shaw (2005) have used the subspecific names of L. infantum infantum and L. infantum chagasi in consideration of previous claims that distinct differences exist between the two organisms based on the kDNA fragment patterns using the restriction endonuclease digestion technique (Jackson et al , 1984; comparison of radioiodinated surface proteins of their promastigotes, and monoclonal antibodies generated against promastigote surfaces (Santoro et al 1986); and comparative radiorespirometry studies (Decker-Jackson & Tang 1982). Recently, Martinez et al (2003) claimed to differentiate L. infantum and L. chagasi by the Random Amplification Polymorphic DNA technique (RAPD) and a single 10-mers long primer.…”

Genetic Diversity Analysis of Chinese Leishmania Isolates and Development of L. donovani Complex Specific Markers by RAPD

“…Meanwhile, through the selection of DNA markers among differentially ampli ed bands, the speci c genetic markers of one species can be developed to species identi cation or assist diagnosis by making a probe. So it proved to be an effective method to obtain genetic markers to develop relevant Leishmania DNA assays [25,26].…”

Molecular Diagnosis of Leishmaniasis, Species Identification and Phylogenetic Analysis

“…RAPD has been used for the investigation of the genomic diversity of L.braziliensis strains [130,131], L.major isolates [132], L.donovani complex [124,133,134] and L.infantum [77,121,135]. Regarding the use of RAPD in species identification, it has been applied for the differentiation between the species L.braziliensis, L.mexicana, L.infantum, L.tropica, L.chagasi, L.amazonensis and L.major [136], the identification and differentiation of Old World species at complex level [137] and recently for the characterization of clinical isolates responsible for kala-azar in India [138]. The main disadvantages of this technique are the need for parasite culture due to the use of non Leishmania specific primers and the poor reproducibility of the assay.…”