Project Lead(s): Claire Mugasa
Issue
Human African trypanosomiasis (HAT) is a highly neglected tropical disease.
About 20 to 30% of HAT patients remain undiagnosed by current parasitological techniques.
In the past decade, only one molecular test – the Loop Mediated Isothermal Amplification – has shown potential for use in remote disease endemic areas.
Solution
The aim of this study was to develop an isothermal helicase-dependant amplification (HDA) assay for the detection of trypanosome DNA, as a proof-of-principle.
HDA is an isothermal test that uses DNA helicase in the presence of binding proteins (SSBs) to generate single stranded DNA (ssDNA) templates by disassociating the double-stranded DNA (dsDNA) for primer hybridization and extension thereafter, using polymerase.
Outcome
HDA was developed using DNA extracted from T. brucei parasites from in-vitro and in-vivo (mice) cultures and based on the 18S ribosomal gene (18S rDNA) primers and the 71-basepair amplicon detected on 2.5% agarose gel.
The analytical sensitivity of this test was 10 parasites per mL of in-vitro culture.
The next step for the research project is to complete development of the prototype test and conduct trials to assess its accuracy.