Related Experiment Video
Updated: Feb 7, 2026

Genetic Variant Detection in the CALR gene using High Resolution Melting Analysis
Published on: August 26, 2020
Amino acid permease 3 (aap3) coding sequence as a target for Leishmania identification and diagnosis of leishmaniases
Karl Erik Müller1, Ricardo Andrade Zampieri2, Juliana Ide Aoki2
1Department of Clinical Science, Faculty of Medicine, University of Bergen, Postboks 7804, 5020, Bergen, Norway. karl.muller@uib.no.
Background:
The leishmaniases comprise a spectrum of clinical manifestations caused by different species of Leishmania. Identification of species is important for diagnosis, treatment and follow-up management. However, there is no gold standard for species identification. High resolution melting analysis (HRM) offers a possibility to differentiate Leishmania species without the need for processing of the PCR-product. The amino acid permease 3 (aap3) gene is an exclusive target for trypanosomatids and is conserved among Leishmania spp., thus it can be a valuable target for an HRM assay for diagnosis of the leishmaniases.
Results:
The HRM dissociation profiles of three amplicons targeting the aap3-coding region allowed the discrimination of L. (Leishmania) donovani, L. (L.) infantum, L. (L.) major, L. (L.) tropica, L. (L.) mexicana, L. (L.) amazonensis, L. (Viannia) braziliensis, L. (V.) guyanensis, L. (V.) lainsoni, L. (V.) naiffi and L. (V.) shawi using DNA from promastigote cultures. The protocol was validated with DNA samples from clinical infection in humans and a cat, naturally infected sand flies, and experimentally infected mice.
Conclusions:
HRM analysis using the aap3 coding sequence as target is a relatively cheap, fast and robust strategy to detect and discriminate Leishmania species from all the endemic regions worldwide. The target and method proved to be useful in clinical, field and experimental samples, thus it could be used as a tool in diagnosis as well as ecological and epidemiological studies.
Related Concept Videos
Amino acids
Amino Acid Catabolism
Amino Acid Biosynthetic Pathways
lncRNA - Long Non-coding RNAs
lncRNA - Long Non-coding RNAs
Phase II Reactions: Sulfation and Conjugation with α-Amino Acids

