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Nuclear DNA sequence specific to Leishmania (Viannia) subgenus: a molecular marker for species identification

Published online by Cambridge University Press:  18 June 2002

L. LUIS ,
A. H. RAMÍREZ ,
R. RAMÍREZ ,
I. D. VÉLEZ  and
A. MENDOZA-LEÓN
L. LUIS
Affiliation:
Laboratorio de Bioquímica y Biología Molecular de Parásitos, Institiuto de Biologia Experimental (I.B.E.), Facultad de Ciencias, Universidad Central de Venezuela, Apartado 47577, Caracas 1041A, Venezuela
A. H. RAMÍREZ
Affiliation:
Laboratorio de Bioquímica y Biología Molecular de Parásitos, Institiuto de Biologia Experimental (I.B.E.), Facultad de Ciencias, Universidad Central de Venezuela, Apartado 47577, Caracas 1041A, Venezuela
R. RAMÍREZ
Affiliation:
Laboratorio de Bioquímica y Biología Molecular de Parásitos, Institiuto de Biologia Experimental (I.B.E.), Facultad de Ciencias, Universidad Central de Venezuela, Apartado 47577, Caracas 1041A, Venezuela Programa de Estudio y Control de Enfermedades Tropicales (PECET), Universidad de Antioquia, Medellin, Colombia
I. D. VÉLEZ
Affiliation:
Programa de Estudio y Control de Enfermedades Tropicales (PECET), Universidad de Antioquia, Medellin, Colombia
A. MENDOZA-LEÓN
Affiliation:
Laboratorio de Bioquímica y Biología Molecular de Parásitos, Institiuto de Biologia Experimental (I.B.E.), Facultad de Ciencias, Universidad Central de Venezuela, Apartado 47577, Caracas 1041A, Venezuela
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Abstract

As shown by RFLP analysis, there is a high variability in the β-tubulin gene region of Leishmania sp. Such variability has been used in the identification of these parasites, establishing differences between subgenera of New World Leishmania. We have found a region of 500 bp (β500) upstream of the coding region of the β-tubulin gene that is present in all strains tested belonging to the L. (Viannia) subgenus. This region apparently is a repetitive sequence and we have shown that it is specific to the Leishmania (Viannia) subgenus. This sequence has no homology with the genomic DNA isolated from either the species belonging to the L. (Leishmania) subgenus or other Kinetoplastida, such as Trypanosoma cruzi, T. brucei, Leptomonas samueli, or Crithidia fasciculata. The β500 sequence showed sufficient variation to be used as a molecular marker in the identification of parasites. We established inter- and intrasubgenus differentiation and were able to discriminate at the species level in the Vianna subgenus. A PCR assay confirmed the specificity of the β500 sequence.

Keywords

LeishmaniaViannia subgenuspolymorphismrepetitive DNAspecies identification
Type
Research Article
Information
Parasitology , Volume 122 , Issue 4 , April 2001 , pp. 405 - 414
Copyright
© 2001 Cambridge University Press

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