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Disease surveillance can be carried out by detecting\n                      <jats:italic>Trypanosoma<\/jats:italic>\n                      DNA in tsetse, also known as molecular xenomonitoring. Whilst molecular methods can increase the efficiency and sensitivity of pathogen detection, trained staff and a well-equipped laboratory are required. In many cases, DNA extraction and screening is outsourced to a central laboratory in a major city either in-country or abroad, far removed from original tsetse collection sites. This increases results turnaround time, incurs transportation costs, and can lead to sample loss or damage.\n                    <\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec id=\"sec002\">\n                    <jats:title>Methodology\/Principle Findings<\/jats:title>\n                    <jats:p>\n                      We set out to develop, optimise and trial methods for tsetse xenomonitoring in a low-resource point-of-need setting. A low-cost protocol was developed consisting of rapid alkali-based DNA extraction and\n                      <jats:italic>Trypanosoma<\/jats:italic>\n                      detection qPCR assays using air-dryable reagent mixes. A minimally-equipped laboratory was established in a field station in Arua, Uganda. Following a training workshop, three entomology technicians carried out screening on 286 tsetse collected over a nine-week study period. The technicians consistently extracted high quality DNA (98% success rate) and were able to successfully detect\n                      <jats:italic>T. brucei sensu lato<\/jats:italic>\n                      in 4.3% (95% confidence interval (CI) [2.23 - 7.37]),\n                      <jats:italic>T. congolense<\/jats:italic>\n                      in 3.6% (95% CI [1.73 \u2013 6.47]) and\n                      <jats:italic>T. vivax<\/jats:italic>\n                      in 3.9% (95% CI [1.98 \u2013 6.92]) of total tsetse, representing a total\n                      <jats:italic>Trypanosoma sp<\/jats:italic>\n                      detection prevalence of 10.7% (95% CI [9.6 \u2013 11.8]).\n                    <\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec id=\"sec003\">\n                    <jats:title>Conclusions\/Significance<\/jats:title>\n                    <jats:p>\n                      This study demonstrated that sensitive molecular xenomonitoring of HAT and AAT pathogens can be carried out without the need for cold-chain storage or high-powered equipment. Further improvements to the system might be achieved by modifying the DNA extraction protocol to enable high-throughput or pooled samples, increasing the sensitivity of the\n                      <jats:italic>T. b. gambiense<\/jats:italic>\n                      DNA detection assay and exploring more sustainable power sources.\n                    <\/jats:p>\n                  <\/jats:sec>","DOI":"10.1371\/journal.pntd.0013706","type":"journal-article","created":{"date-parts":[[2026,3,23]],"date-time":"2026-03-23T17:28:03Z","timestamp":1774286883000},"page":"e0013706","update-policy":"https:\/\/doi.org\/10.1371\/journal.pntd.corrections_policy","source":"Crossref","is-referenced-by-count":0,"title":["Development and pilot application of a point-of-need molecular xenomonitoring protocol for tsetse (Glossina sp.) in a low-resource setting"],"prefix":"10.1371","volume":"20","author":[{"ORCID":"https:\/\/orcid.org\/0000-0003-4141-8625","authenticated-orcid":true,"given":"Isabel","family":"Saldanha","sequence":"first","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Edward","family":"Aziku","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Alex T.","family":"Trima","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Victor","family":"Drapari","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Gala","family":"Garrod","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Henry","family":"Ombanya","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Inaki","family":"Tirados","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Martha","family":"Betson","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Albert","family":"Mugenyi","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Sophie","family":"Dunkley","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Stephen J.","family":"Torr","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Andrew","family":"Hope","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Lucas J.","family":"Cunningham","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"340","published-online":{"date-parts":[[2026,3,23]]},"reference":[{"key":"pntd.0013706.ref001","doi-asserted-by":"crossref","first-page":"284","DOI":"10.1186\/s13071-015-0898-y","article-title":"Developing a continental atlas of the distribution and trypanosomal infection of tsetse flies (Glossina species)","volume":"8","author":"G Cecchi","year":"2015","journal-title":"Parasit Vectors"},{"issue":"7527","key":"pntd.0013706.ref002","doi-asserted-by":"crossref","first-page":"1238","DOI":"10.1136\/bmj.331.7527.1238","article-title":"Sleeping sickness in Uganda: a thin line between two fatal diseases","volume":"331","author":"K Picozzi","year":"2005","journal-title":"BMJ"},{"issue":"5","key":"pntd.0013706.ref003","doi-asserted-by":"crossref","first-page":"384","DOI":"10.1097\/QCO.0000000000000860","article-title":"Gambiense human African trypanosomiasis: the bumpy road to elimination","volume":"35","author":"E Hasker","year":"2022","journal-title":"Curr Opin Infect Dis"},{"key":"pntd.0013706.ref004","unstructured":"World Health Organisation. 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