Intergenic region typing (IRT): A rapid molecular approach to the characterization and evolution of Leishmania

Elisa Cupolillo, Gabriel Grimaldi, Hooman Momen, Stephen M. Beverley

Research output: Contribution to journalArticle

180 Scopus citations

Abstract

In the New World, Leishmania of the Viannia subgenus cause both cutaneous and mucocutaneous disease. These parasites show considerable intra-species genetic diversity and variation, which complicates taxonomic classification and epidemiology. We have used the variability of the transcribed noncoding regions between the small and large subunit rRNA genes to examine relationships in this group. In a method termed intergenic region typing (IRT), PCR amplification products were obtained for the rapidly evolving 1 - 1.2-kb internal transcribed spacers (ITS) between the SSU and LSU rRNAs, from 50 parasites isolated from different hosts and geographic areas. Amplified DNAs were digested with 10 different enzymes, and fragment patterns compared after acrylamide gel eletrophoresis. High levels of intra- and inter-specific variation were observed, and quantitative similarity comparisons were used to associate different lineages. A complex evolutionary tree was obtained. Some species formed tight clusters (L. equatorensis, L. panamensis, L. guyanensis, L. shawi), while L. braziliensis was highly polymorphic and L. naiffi showed intraspecific distances comparable to the largest obtained within all Viannia. L. colombiensis, L. equatorensis and L. lainsoni clearly represent distinct lineages. Good agreement was obtained with molecular trees based upon isoenzyme or mini-exon repeat sequence comparisons. Overall, IRT appears to be a superb method for epidemiological and taxonomic studies of Leishmania, being sensitive, rapid and quantitative while simultaneously revealing considerable molecular diversity. IRT could also be applied to other nonconserved intergenic regions, including those separating protein-coding genes.

Original languageEnglish
Pages (from-to)145-155
Number of pages11
JournalMolecular and Biochemical Parasitology
Volume73
Issue number1-2
DOIs
StatePublished - Jul 1995
Externally publishedYes

Keywords

  • Epidemiology
  • Ribosomal DNA
  • Taxonomy
  • Transcribed intergenic region
  • Viannia

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