{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,11,2]],"date-time":"2026-11-02T23:12:21Z","timestamp":1793661141958,"version":"4.5.1"},"reference-count":36,"publisher":"MDPI AG","issue":"2","license":[{"start":{"date-parts":[[2021,4,30]],"date-time":"2021-04-30T00:00:00Z","timestamp":1619740800000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["TropicalMed"],"abstract":"<jats:p>African animal trypanosomiasis (AAT) control programs rely on active case detection through the screening of animals reared in disease endemic areas. This study compared the application of the polymerase chain reaction (PCR) and microscopy in the detection of trypanosomes in cattle blood in Mambwe, a rural district in eastern Zambia. Blood samples were collected from 227 cattle and tested for infection with trypanosomes using microscopy and Ribosomal RNA Internal Transcribed Spacers (ITS)-PCR. Microscopy on the buffy coat detected 17 cases, whilst thin and thick smears detected 26 cases and 28 cases, respectively. In total, microscopy detected 40 cases. ITS-PCR-filter paper (FP) on blood spots stored on FP detected 47 cases, and ITS-PCR-FTA on blood spots stored on Whatman FTA Classic cards detected 83 cases. Using microscopy as the gold standard, ITS-PCR-FTA had a better specificity (SP) and sensitivity (SE) (SP = 72.2%; SE = 77.5%; kappa = 0.35) than ITS-PCR-FP (SP = 88%; SE = 60%; kappa = 0.45). The prevalence of Trypanosoma brucei s.l. was higher on ITS-PCR-FTA (19\/227) than on ITS-PCR-FP (0\/227). Our results illustrate the complexities around trypanosomiasis surveillance in rural Africa and provide evidence of the impact that field conditions and staff training can have on diagnostic results, which in turn impact the success of tsetse and trypanosomiasis control programs in the region.<\/jats:p>","DOI":"10.3390\/tropicalmed6020068","type":"journal-article","created":{"date-parts":[[2021,4,30]],"date-time":"2021-04-30T10:55:04Z","timestamp":1619780104000},"page":"68","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":16,"title":["Challenges in the Diagnostic Performance of Parasitological and Molecular Tests in the Surveillance of African Trypanosomiasis in Eastern Zambia"],"prefix":"10.3390","volume":"6","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-3611-5016","authenticated-orcid":false,"given":"Gloria M.","family":"Mulenga","sequence":"first","affiliation":[{"name":"College of Public Health Medical and Veterinary Services, James Cook University, Townsville, QLD 4814, Australia"},{"name":"Department of Veterinary Services, Ministry of Fisheries and Livestock, Lusaka 10101, Zambia"},{"name":"Department of Veterinary Services, Kakumbi Tsetse and Trypanosomiasis Research Station, P.O. Box 70, Mfuwe 10101, Zambia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Boniface","family":"Namangala","sequence":"additional","affiliation":[{"name":"Institute of Distance Learning, The University of Zambia, Lusaka 10101, Zambia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Kalinga","family":"Chilongo","sequence":"additional","affiliation":[{"name":"Department of Veterinary Services, Ministry of Fisheries and Livestock, Lusaka 10101, Zambia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Chrisborn","family":"Mubamba","sequence":"additional","affiliation":[{"name":"Department of Veterinary Services, Ministry of Fisheries and Livestock, Lusaka 10101, Zambia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Kyoko","family":"Hayashida","sequence":"additional","affiliation":[{"name":"Research Centre for Zoonosis Control, Hokkaido University, Sapporo 0010020, Japan"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-3533-1515","authenticated-orcid":false,"given":"Lars","family":"Henning","sequence":"additional","affiliation":[{"name":"College of Public Health Medical and Veterinary Services, James Cook University, Townsville, QLD 4814, Australia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Bruce","family":"Gummow","sequence":"additional","affiliation":[{"name":"College of Public Health Medical and Veterinary Services, James Cook University, Townsville, QLD 4814, Australia"},{"name":"Faculty of Veterinary Science, University of Pretoria, Pretoria 0028, South Africa"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2021,4,30]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"2017","DOI":"10.1016\/S0140-6736(01)07096-9","article-title":"Identification of human-infective trypanosomes in animal reservoir of sleeping sickness in Uganda by means of serum-resistance-associated (SRA) gene","volume":"358","author":"Welburn","year":"2001","journal-title":"Lancet"},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"231","DOI":"10.1016\/j.vetpar.2007.04.005","article-title":"The comparative role of cattle, goats and pigs in epidemiology of livestock trypanosomiasis on plateau of eastern Zambia","volume":"147","author":"Simukoko","year":"2007","journal-title":"Vet. Parasitol."},{"key":"ref_3","unstructured":"(2021, April 29). PAAT\u2014Programme Against African Trypanosomiasis. Available online: http:\/\/www.fao.org\/paat\/the-programme\/our-structure\/es\/."},{"key":"ref_4","unstructured":"FAO (Food and Agriculture Organization of the United Nations) (2018). WHO and FAO Step up towards the Elimination of Human African Trypanosomiasis to Ensure Health and Food Security in Africa, FAO."},{"key":"ref_5","doi-asserted-by":"crossref","unstructured":"Katsidzira, L., and Fana, G.T. (2010). Pitfalls in the diagnosis of Trypanosomiasis in low endemic countries: A case Report. PLoS Negl. Trop. Dis., 4.","DOI":"10.1371\/journal.pntd.0000823"},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"313","DOI":"10.4269\/ajtmh.2004.71.313","article-title":"Chappuis F, Stivanello E, Adams K, Ki Card agglutination test for trypanosomisis end-dilution titer and cerebrospinal fluid cell count as predictors of human African trypanosomiasis (T. b. r) among serologically susp","volume":"71","author":"Chappuis","year":"2004","journal-title":"Am. J. Trop. Med. Hyg."},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"82","DOI":"10.1186\/1756-3305-3-82","article-title":"Constraints to estimating the prevalence of trypanosome infections in East African zebu cattle","volume":"3","author":"Cox","year":"2010","journal-title":"Parasites Vectors"},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"840","DOI":"10.1111\/j.1365-3156.2005.01470.x","article-title":"Quantifying the level of under detection of T. b. r. sleeping sickness cases","volume":"10","author":"Odiit","year":"2005","journal-title":"Trop. Med. Int. Health"},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"1","DOI":"10.1186\/1756-3305-1-46","article-title":"Comparative evaluation of three PCR base diagnostic assays for the detection of pathogenic trypanosomes in cattle blood","volume":"1","author":"Thumbi","year":"2008","journal-title":"Parasites Vectors"},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"253","DOI":"10.1016\/j.vetpar.2014.04.014","article-title":"Detection of African animal trypanosomes: The haematocrit centrifugation technique compared to PCR with samples stored on filter paper or in DNA protecting buffer","volume":"203","author":"Moti","year":"2014","journal-title":"Vet. Parasitol."},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"24","DOI":"10.1016\/j.exppara.2005.03.014","article-title":"A PCR based assay for detection and differentiation of African trypanosome species in blood","volume":"22","author":"Cox","year":"2005","journal-title":"Exp. Parasitol."},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"610","DOI":"10.1016\/S0020-7519(01)00161-8","article-title":"Detection and identification of Trypanosoma of African livestock through a single PCR based on internal transcribed spacer 1 of rDNA","volume":"31","author":"Desquesnes","year":"2001","journal-title":"Int. J. Parasitol."},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"1819","DOI":"10.1292\/jvms.16-0240","article-title":"A PCR-based survey of animal African trypanosomosis and selected piroplasm parasites of cattle and goats in Zambia","volume":"78","author":"Musinguzi","year":"2017","journal-title":"J. Vet. Med. Sci."},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"589","DOI":"10.1016\/j.ijpara.2007.09.006","article-title":"African trypanosomiasis: Sensitive and rapid detection of the sub-genus Trypanozoon by loop-mediated isothermal amplification (LAMP) of parasite DNA","volume":"38","author":"Njiru","year":"2008","journal-title":"Int. J. Parasitol."},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"349","DOI":"10.1016\/S0001-706X(02)00029-3","article-title":"Comparison of different DNA preparation protocols for PCR diagnosis of HAT in Cote d\u2019ivoire","volume":"82","author":"Solano","year":"2002","journal-title":"Acta. Trop."},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"186","DOI":"10.1007\/s00436-004-1267-5","article-title":"The use of ITS1 rDNA PCR in detecting pathogenic African trypanosomes","volume":"95","author":"Njiru","year":"2005","journal-title":"Parasitol. Res."},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"555","DOI":"10.1186\/s13071-015-1173-y","article-title":"Studies of trypanosomiasis in the Luangwa valley, north-eastern Zambia","volume":"8","author":"Laohasinnarong","year":"2015","journal-title":"Parasites Vectors"},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"68","DOI":"10.1186\/1756-3305-4-68","article-title":"The best practice for preparation of samples from FTA(R)cards for diagnosis of blood borne infections using African trypanosomes as a model system","volume":"4","author":"Ahmed","year":"2011","journal-title":"Parasites Vectors"},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"104","DOI":"10.1186\/s13071-020-3984-8","article-title":"The buffy coat method: A tool for detection of blood parasites without staining procedures","volume":"13","author":"Chagas","year":"2020","journal-title":"Parasites Vectors"},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"638","DOI":"10.1186\/s13071-015-1260-0","article-title":"Prevalence of pathogenic trypanosomes in anaemic cattle from trypanosomosis challenged areas of Itezhi-tezhi district in central Zambia","volume":"8","author":"Mbewe","year":"2015","journal-title":"Parasites Vectors"},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"288","DOI":"10.1016\/j.prevetmed.2008.05.002","article-title":"Evaluating the use of packed cell volume as an indicator of trypanosomal infections in cattle in eastern Zambia","volume":"87","author":"Marcotty","year":"2008","journal-title":"Prev. Vet. Med."},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"1132","DOI":"10.4269\/ajtmh.2007.76.1132","article-title":"Use of multiple displacement amplification to increase the detection and genotyping of trypanosoma species samples immobilized on FTA filters","volume":"76","author":"Morrison","year":"2007","journal-title":"Am. J. Trop. Med. Hyg."},{"key":"ref_23","doi-asserted-by":"crossref","unstructured":"Gaithuma, A.K., Yamagishi, J., Martinelli, A., Hayashida, K., Kawai, N., Marsela, M., and Sugimoto, C. (2019). A single test approach for accurate and sensitive detection and taxonomic characterization of Trypanosomes by comprehensive analysis of internal transcribed spacer 1 amplicons. PLoS. Negl. Trop. Dis., 13.","DOI":"10.1371\/journal.pntd.0006842"},{"key":"ref_24","doi-asserted-by":"crossref","unstructured":"Ahmed, H.A., Picozzi, K., Welburn, S.C., and MacLeod, E.T. (2013). A comparative evaluation of PCR- based methods for species- specific determination of African animal trypanosomes in Ugandan cattle. Parasites Vectors.","DOI":"10.1186\/1756-3305-6-316"},{"key":"ref_25","doi-asserted-by":"crossref","unstructured":"Sawitri, D.H., Wardhana, A.H., Dewi, D.A., Ekawasti, F., and Widjaja, E. (2016, January 10). Application of Dried Blood Sample on Whatman Filter Paper for Detection of Trypanosoma evansi from Cattle in Central Kalimantan by Internal Trascriber Spacer-1 Polymerase Chain Reaction. Proceedings of the International Seminar on Livestock Production and Veterinary Technology, Bali, Indonesia.","DOI":"10.14334\/Proc.Intsem.LPVT-2016-p.191-197"},{"key":"ref_26","first-page":"S83","article-title":"Sensitivity, Specificity, Receiver-Operating Characteristic (ROC) Curves and Likelihood Ratios: Communicating the Performance of Diagnostic Test","volume":"29","author":"Florkwoski","year":"2008","journal-title":"Clin. Biochem. Rev."},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"174","DOI":"10.1016\/j.vetpar.2015.09.001","article-title":"Detection of Trypanosoma vivax using PCR and LAMP during aparasitemic periods","volume":"214","author":"Cadioli","year":"2015","journal-title":"Vet. Parasitol."},{"key":"ref_28","doi-asserted-by":"crossref","unstructured":"de Clare Bronsvoort, B.M., von Wissmann, B., Fevre, E.M., Handel, I.G., Picozzi, K., and Welburn, S.C. (2010). No gold standard estimation of the sensitivity and specificity of two molecular diagnostic protocols for Trypanosoma brucei spp. in Western Kenya. PLoS ONE, 5.","DOI":"10.1371\/journal.pone.0008628"},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"40","DOI":"10.1016\/0035-9203(80)90009-7","article-title":"Miniature anion-exchange\/centrifugation technique for the diagnosis of microfilaraemia in the field","volume":"74","author":"Lumsden","year":"1980","journal-title":"Trans. R. Soc. Soc. Trop. Med. Hyg."},{"key":"ref_30","unstructured":"Despommier, G., and Hotez, K. (2005). The Protozoa. Parasitic Diseases, Apple Trees Productions."},{"key":"ref_31","doi-asserted-by":"crossref","unstructured":"Muhanguzi, D., Mugenyi, A., Bigirwa, G., Kamusiime, M., Kitibwa, A., Akurut, G.G., Ochwo, S., Amanyire, W., Okech, S.G., and Hattendorf, J. (2017). African animal trypanosomiasis as a constraint to livestock health and production in Karamoja region: A detailed qualitative and quantitative assessment. BMC Vet. Res., 13.","DOI":"10.1186\/s12917-017-1285-z"},{"key":"ref_32","doi-asserted-by":"crossref","first-page":"51","DOI":"10.1016\/j.rvsc.2010.04.021","article-title":"Bovine trypanosomiasis risk in an endemic area on the eastern plateau of Zambia","volume":"90","author":"Simukoko","year":"2011","journal-title":"Res. Vet. Sci."},{"key":"ref_33","doi-asserted-by":"crossref","first-page":"1162","DOI":"10.1017\/S0031182017000440","article-title":"Characterization of tabanid flies (Diptera: Tabanidae) in South Africa and Zambia and detection of protozoan parasites they are harbouring","volume":"144","author":"Taioe","year":"2017","journal-title":"Parasitology"},{"key":"ref_34","doi-asserted-by":"crossref","first-page":"116","DOI":"10.4269\/ajtmh.13-0078","article-title":"Preliminary investigation of trypanosomosis in exotic dog breeds from Zambia\u2019s Luangwa and Zambezi valleys using LAMP","volume":"89","author":"Namangala","year":"2013","journal-title":"Am. J. Trop. Med. Hyg."},{"key":"ref_35","doi-asserted-by":"crossref","first-page":"510","DOI":"10.1186\/s13071-015-1125-6","article-title":"The role of domestic animals in the epidemiology of human African trypanosomiasis in Ngorongoro conservation area, Tanzania","volume":"8","author":"Ruiz","year":"2015","journal-title":"Parasites Vectors"},{"key":"ref_36","doi-asserted-by":"crossref","first-page":"281","DOI":"10.1186\/1756-3305-6-281","article-title":"Cattle movements and trypanosomes: Restocking efforts and the spread of Trypanosoma brucei rhodesiense sleeping sickness in post-conflict Uganda","volume":"6","author":"Selby","year":"2013","journal-title":"Parasites Vectors"}],"container-title":["Tropical Medicine and Infectious Disease"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/2414-6366\/6\/2\/68\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,11]],"date-time":"2025-10-11T05:56:13Z","timestamp":1760162173000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/2414-6366\/6\/2\/68"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2021,4,30]]},"references-count":36,"journal-issue":{"issue":"2","published-online":{"date-parts":[[2021,6]]}},"alternative-id":["tropicalmed6020068"],"URL":"https:\/\/doi.org\/10.3390\/tropicalmed6020068","relation":{},"ISSN":["2414-6366"],"issn-type":[{"value":"2414-6366","type":"electronic"}],"subject":[],"published":{"date-parts":[[2021,4,30]]}}}