Development of multiplex serological assay for the detection of human African trypanosomiasis |
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| Institution: | 1. Nagasaki University Institute of Tropical Medicine (NUITM), - Kenya Medical Research Institute (KEMRI) Project, Box 19993-00202 Nairobi, Kenya;2. Centre for Infectious and Parasitic Diseases Control Research, Kenya Medical Research Institute (KEMRI), Box 3-50400 Busia, Kenya;3. Department of Eco-epidemiology, Institute of Tropical Medicine, Nagasaki University (NUITM), 1-12-24 Sakamaoto, Nagasaki 852-8523, Japan;4. Production Department, Kenya Medical Research Institute (KEMRI), Box 54840-00200, Nairobi, Kenya;5. Department of Infection and Immunology, Aichi Medical University School of Medicine, 1-1 Yazakokarimata, Nagakute, Aichi Prefecture 480-1195, Japan;6. Department of Microbiology, Rajshahi Medical College, Laxmipur, 6000 Rajshahi, Bangladesh;7. Department of Parasitology, Institute of Tropical Medicine, Nagasaki University (NUITM), 1-12-24 Sakamaoto, Nagasaki 852-8523, Japan;8. Department of Immunogenetics, Institute of Tropical Medicine, Nagasaki University (NUITM), 1-12-24 Sakamaoto, Nagasaki 852-8523, Japan;9. Consortium for National Health Research (CNHR), Box 29832-00202 Nairobi, Kenya;10. Graduate School of International Health Development, Nagasaki University, 1-12-24 Sakamaoto, Nagasaki 852-8523, Japan |
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| Abstract: | Human African trypanosomiasis (HAT) is a disease caused by Kinetoplastid infection. Serological tests are useful for epidemiological surveillance. The aim of this study was to develop a multiplex serological assay for HAT to assess the diagnostic value of selected HAT antigens for sero-epidemiological surveillance.We cloned loci encoding eight antigens from Trypanosoma brucei gambiense, expressed the genes in bacterial systems, and purified the resulting proteins. Antigens were subjected to Luminex multiplex assays using sera from HAT and VL patients to assess the antigens' immunodiagnostic potential. Among T. b. gambiense antigens, the 64-kDa and 65-kDa invariant surface glycoproteins (ISGs) and flagellar calcium binding protein (FCaBP) had high sensitivity for sera from T. b. gambiense patients, yielding AUC values of 0.871, 0.737 and 0.858 respectively in receiver operating characteristics (ROC) analysis. The ISG64, ISG65, and FCaBP antigens were partially cross-reactive to sera from Trypanosoma brucei rhodesiense patients. The GM6 antigen was cross-reactive to sera from T. b. rhodesiense patients as well as to sera from VL patients. Furthermore, heterogeneous antibody responses to each individual HAT antigen were observed. Testing for multiple HAT antigens in the same panel allowed specific and sensitive detection. Our results demonstrate the utility of applying multiplex assays for development and evaluation of HAT antigens for use in sero-epidemiological surveillance. |
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