Original Research

Comparative evaluation of dry and liquid RIME LAMP in detecting trypanosomes in dead tsetse flies

Peter Nambala, Janelisa Musaya, Kyoko Hayashida, Emmanuel Maganga, Edward Senga, Kelita Kamoto, John Chisi, Chihiro Sugimoto
Onderstepoort Journal of Veterinary Research | Vol 85, No 1 | a1543 | DOI: https://doi.org/10.4102/ojvr.v85i1.1543 | © 2018 Peter Nambala, Janelisa Musaya, Kyoko Hayashida, Emmanuel Maganga, Edward Senga, Kelita Kamoto, John Chisi, Chihiro Sugimoto | This work is licensed under CC Attribution-NoDerivatives 4.0
Submitted: 11 September 2017 | Published: 03 October 2018

About the author(s)

Peter Nambala, Department of Basic Medical Sciences, University of Malawi, Malawi
Janelisa Musaya, Department of Pathology, University of Malawi, Malawi
Kyoko Hayashida, Research Centre for Zoonosis Control, Hokkaido University, Japan
Emmanuel Maganga, Mikolongwe Veterinary College of Agriculture and Food Security, Limbe, Malawi
Edward Senga, Department of Basic Medical Sciences, University of Malawi, Malawi
Kelita Kamoto, Department of Basic Medical Sciences, University of Malawi, Malawi
John Chisi, Department of Basic Medical Sciences, University of Malawi, Malawi
Chihiro Sugimoto, Research Centre for Zoonosis Control, Hokkaido University, Japan


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Abstract

Xenomonitoring is an important approach in assessing the progress of trypanosomiasis control as well as in estimating the endemicity of trypanosomes in affected areas. One of the major challenges in this approach is the unavailability of sensitive and easy to use xenomonitoring tools that can be used in the remote areas where the disease occurs. One tool that has been used successfully in detecting the parasites in tsetse flies is the repetitive insertion mobile element loop-mediated isothermal amplification (RIME LAMP). This tool has recently been modified from the liquid form to dry form for use in remote areas; however, uptake for use in the field has been slow. Field-collected tsetse flies were used to evaluate the performance of dry RIME LAMP over the conventional liquid RIME LAMP. All the samples were also subjected to internal transcribed spacer 1 (ITS1) ribosomal deoxyribonucleic acid (DNA) polymerase chain reaction (PCR) as a standard. ITS1-PCR-positive samples were further sequenced for confirmation of the species. A total of 86 wild tsetse flies were left to dry at room temperature for 3 months and DNA was extracted subsequently. All 86 flies were Glossina morsitans morsitans. From these, dry RIME LAMP detected 16.3% while liquid RIME LAMP detected 11.6% as infected with trypanosomes. Ten positive samples on ITS1-PCR were sequenced and all were shown to be trypanosomes. The use of dry RIME LAMP in the field for xenomonitoring of trypanosomes in tsetse flies will greatly contribute towards control of this neglected tropical disease as it provides the cheapest, fastest and simplest way to estimate possible human infective trypanosome infection rates in the tsetse fly vectors.

Keywords

Dry RIME LAMP; Tsetse flies; Trypanosmes

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