Instrument-Free Point-of-Care Diagnostic for Leishmania Parasites

Area of Science:

• Parasitology
• Molecular Diagnostics
• Biotechnology

Background:

• Leishmaniasis is a severe parasitic disease affecting 1.6 million annually, with potential for life-altering complications.
• Accurate species diagnosis is vital for effective leishmaniasis treatment and patient follow-up.
• Current PCR diagnostics are slow, costly, and unsuitable for resource-limited areas.

Purpose of the Study:

• To develop a simple, affordable, and instrument-free diagnostic tool for Leishmania DNA detection.
• To enable sensitive parasite detection and species identification in both high-tech and field settings.
• To address the limitations of existing diagnostic methods for leishmaniasis.

Main Methods:

• Computational biology was used to design RPA oligos and CRISPR guides for Leishmania detection.
• Methods included parasite lysis, RPA amplification, and fluorescent CRISPR crRNA detection.
• Specific identification of *L. (V.) panamensis* was targeted as a predictor of mucocutaneous disease.

Main Results:

• Demonstrated detection of single-digit parasites with high species specificity.
• Successfully developed a proof-of-concept for a point-of-care diagnostic.
• Achieved quantifiable fluorescent signals over negative controls in successive assays.

Conclusions:

• The study provides a foundation for a three-track assay (all Leishmania, specific species, human control).
• The assay will be implemented in a Funnel Adapted Sensing Tube (FAST) device.
• The FAST device aims to be single-use, instrument-free, and affordable for widespread use.