Apr 21, 2020

Identification of Trypanosoma brucei gambiense and T. b. rhodesiense in vectors using multiplexed high-resolution melt analysis.

BioRxiv : the Preprint Server for Biology
Gala GarrodL. J. Cunningham


Background Human African Trypanosomiasis (HAT) is a potentially fatal parasitic infection caused by the trypanosome sub-species Trypanosoma brucei gambiense and T. b. rhodesiense transmitted by tsetse flies. Currently, global HAT case numbers are reaching less than 1 case per 10,000 people in many disease foci. As such, there is a need for simple screening tools and strategies to replace active screening of the human population which can be maintained post-elimination for Gambian HAT and long-term Rhodesian HAT. Here we describe the development of a novel high-resolution melt assay for the xenomonitoring of Trypanosoma brucei gambiense and T. b. rhodesiense in tsetse. Methods Primers for T. b. rhodesiense and T. b. gambiense were designed to target species-specific single copy genes. An additional primer set was included in the multiplex to determine if samples have sufficient genomic material for detecting low copy number targets. The assay was evaluated on 96 wild-caught tsetse previously identified to be positive for T. brucei s. l. of which two were infected with T. b. rhodesiense. Results The assay was found to be highly specific with no cross-reactivity with non-target trypanosome species and the assay limit of detection ...Continue Reading

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Neuronal Circuitry
Cellular Component Organization
Gene Clusters
Drosophila melanogaster

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