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Sensitive Molecular Diagnostics for Cutaneous Leishmaniasis
Orli Sagi1, Anat Berkowitz1,2, Shlomi Codish3
1Clinical Parasitology.
Insights
A new multiplex qPCR test accurately diagnoses cutaneous leishmaniasis (CL) and identifies Leishmania species from lesion swabs. This rapid diagnostic tool aids in managing CL caused by Old World and New World Leishmania strains.
Area of Science:
- Medical Parasitology
- Molecular Diagnostics
- Infectious Diseases
Background:
- Cutaneous leishmaniasis (CL) diagnosis and species identification are crucial for effective disease management.
- In Israel, CL is primarily caused by Leishmania major and Leishmania tropica.
- Accurate and rapid diagnostic methods are needed for timely treatment.
Purpose of the Study:
- To develop and validate a point-of-care diagnostic test for Leishmania species.
- To enable rapid and accurate identification of Leishmania parasites in clinical samples.
- To differentiate between Old World and New World Leishmania species.
Main Methods:
- Establishment of a lesion-swabbing technique for sample collection.
- Development of a sensitive multiplex real-time PCR (qPCR) assay.
- Utilized three probes for general Leishmania, L. major, and L. tropica detection.
Main Results:
- Screened 1783 clinical samples over two years.
- Detected Leishmania species in 1086 individuals (1008 L. major, 70 L. tropica).
- Identified L. braziliensis and L. infantum/donovani in 8 samples using a secondary multiplex qPCR.
Conclusions:
- The multiplex qPCR test provides accurate diagnostics for Leishmania infections.
- Enabled subtyping of dominant Leishmania strains in the region.
- Facilitated better treatment strategies for CL patients, including travelers.
Background:
Rapid diagnosis of cutaneous leishmaniasis (CL) and identification of Leishmania species is highly important for the disease management. In Israel, CL is caused mainly by Leishmania major and Leishmania tropica species.
Methods:
We established an easy to handle point of care lesion-swabbing, combined with a highly sensitive multiplex real time PCR (multiplex qPCR) for accurate and rapid diagnosis of Leishmania species.
Results:
Using three probes: one general for: Leishmania species, and two specific for L major, and L tropica, we screened 1783 clinical samples collected during two years. Leishmania species was found in 1086 individuals, 1008 L major, and 70 L tropica. Eight samples positive for Leishmania species only, were further tested using a second set of multiplex qPCR developed, and were found positive for Leishmania braziliensis and Leishmania infantum/donovani (2 and 6 samples, concomitantly).
Conclusions:
Taken together, the test enabled diagnostics and better treatment of Leishmania infections from the Old World (1078 samples) and the New World (8 samples), and the subtyping of the dominant strains in the region, as well as in returning travelers'.
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