Related Experiment Video
Updated: Jun 23, 2025
Development of Leishmania Species Strains with Constitutive Expression of eGFP
Published on: April 21, 2023
Rapid detection, quantification and speciation of Leishmania using real-time PCR and DNA sequencing at the rRNA
Andrea Paun1, Michael E Grigg1
1Laboratory of Parasitic Diseases, Molecular Parasitology Section, National Institutes of Health, Bethesda, MD, USA.
Abstract:
The ability to discriminate infection between closely related Leishmania species within the Viannia species complex, specifically L. braziliensis, L. guyanensis and L. panamensis is critical to inform the clinical diagnosis and determine the most efficacious treatment modality. We designed a nested primer set targeting the rRNA Internal Transcribed Spacer 2 (ITS2), located on Chromosome 27, to distinguish among all human infective Leishmania species. Separate nested and single primer pairs were developed for conventional and quantitative PCR approaches respectively. Species-specific single nucleotide polymorphisms and indels located within the PCR products were identified by Sanger sequencing. This single locus approach provides a sensitive and specific platform to identify the species of Leishmania causing infection.
