title: Typing of four genetic loci discriminates among closely related species of New World Leishmania.
abstract: All New World Leishmania species can cause cutaneous lesions, while only Leishmania (Viannia) braziliensis has been associated with mucosal metastases. Multilocus enzyme electrophoresis (MLEE) is the optimal standard for species identification but is slow and costly. New methods for species identification are needed to ensure proper identification and therapy. The coding regions of four metabolic enzyme markers in the MLEE typing method: mannose phosphate isomerase (MPI), malate dehydrogenase (MDH), glucose-6-phosphate isomerase (GPI), and 6-phosphogluconate dehydrogenase (6PGD), were analysed from seven species of New World Leishmania isolated from patients with either cutaneous or mucosal lesions to identify specific genetic polymorphisms responsible for the phenotypic variations observed in the MLEE typing scheme. We identified species-specific polymorphisms and determined that a combination of sequencing of the mpi and 6pgd genes was sufficient to differentiate among seven closely related species of New World Leishmania and among isolates of L. braziliensis shown previously to have atypical MLEE patterns. When DNA isolated from 10 cutaneous lesion biopsies were evaluated, the sequence typing method was 100% concordant with the published MLEE/monoclonal antibody identification methods. The identification of species-specific polymorphisms can be used to design a DNA-based test with greater discriminatory power that requires shorter identification times. When the causative agent of the disease is L. braziliensis, this method ensures correct species identification, even when the agent is a genetic variant. Proper identification could facilitate adequate treatment, preventing the onset of the disfiguring mucosal form of the disease.
authors.affiliation: Parasitology Program, Naval Medical Research Center Detachment, Lima, Peru.
authors.forename: Pablo
authors.initials: P
authors.lastname: Tsukayama
authors.forename: Carmen
authors.initials: C
authors.lastname: Lucas
authors.forename: David J
authors.initials: DJ
authors.lastname: Bacon
mesh_terms: D000818:Animals; D002489:Central America; D016054:DNA, Protozoan / Q000032:analysis / Q000235:genetics; D056426:Genetic Loci; D014644:Genetic Variation; D018503:Genome, Protozoan; D005956:Glucose-6-Phosphate Isomerase / Q000032:analysis / Q000235:genetics; D006801:Humans; D007891:Leishmania / Q000201:enzymology / Q000235:genetics* / Q000302:isolation & purification*; D016773:Leishmaniasis, Cutaneous / Q000175:diagnosis* / Q000201:enzymology / Q000469:parasitology; D008291:Malate Dehydrogenase / Q000032:analysis / Q000235:genetics; D008359:Mannose-6-Phosphate Isomerase / Q000032:analysis / Q000235:genetics; D008969:Molecular Sequence Data; D010734:Phosphogluconate Dehydrogenase / Q000032:analysis / Q000235:genetics; D020641:Polymorphism, Single Nucleotide; D017422:Sequence Analysis, DNA; D013020:South America; D013045:Species Specificity
chemical_list: D016054:DNA, Protozoan; D008291:Malate Dehydrogenase; D010734:Phosphogluconate Dehydrogenase; D008359:Mannose-6-Phosphate Isomerase; D005956:Glucose-6-Phosphate Isomerase