METHOD AND KIT FOR LEISHMANIA (VIANNIA) DIFFERENTIATION OF LEISHMANIA (LEISHMANIA), CAUSES OF LEISHMANIASIS, BY PCR-RFLP

摘要

Method and kit for the differentiation of Leishmania (Leishmania (Leishmania) Viannia), causing leishmaniasis by PCR - RFLP.The objective of this invention is molecular differentiation of species of parasites Leishmania (L. Viannia)(Leishmania), more particularly, between the subgenera Leishmania braziliensis by species (Viannia) and / or L. (v.) guyanensis L. (Leishmania) amazonensis by PCR - RFLP.A first implementation of this invention relates to a method for differentiation between these two species of Leishmania.Through the amplification by PCR of the region kept the minicu00edrculo of kDNA Leishmania.Followed by cleavage of the PCR products with a restriction enzyme specific and analysis of fragments generated by appropriate technique, such as, for example, by RFLP.In the second place, the invention is directed to diagnostic kit to be used in the method of differentiation between the species of parasites Leishmania (L. Viannia)(Leishmania), more particularly, L. (v.) braziliensis by and / or L. (v.) guyanensis of L. (L.The present invention) amazonensis comprising all the reagents required for the completion of the PCR technique, namely: specific primers.Nucleotides and buffer solution suitable for amplification by PCR, Taq polymerase, and appropriate restriction enzyme buffer, the DNA of strains of Leishmania (Viannia reference species) and L.(Leishmania), more particularly, L. (v.) Leishmania braziliensis by, (v.) guyanensis and L. (L.) amazonensis, to serve as a standard and a protocol and manual for instructing the user.