Nzou Samson Muuo, Fujii Yoshito, Miura Masashi, Mwau Matilu, Mwangi Anne Wanjiru, Itoh Makoto, Salam Md Abdus, Hamano Shinjiro, Hirayama Kenji, Kaneko Satoshi
Nagasaki University Institute of Tropical Medicine (NUITM), - Kenya Medical Research Institute (KEMRI) Project, Box 19993-00202 Nairobi, Kenya; Centre for Infectious and Parasitic Diseases Control Research, Kenya Medical Research Institute (KEMRI), Box 3-50400 Busia, Kenya.
Department of Eco-epidemiology, Institute of Tropical Medicine, Nagasaki University (NUITM), 1-12-24 Sakamaoto, Nagasaki 852-8523, Japan.
Parasitol Int. 2016 Apr;65(2):121-7. doi: 10.1016/j.parint.2015.10.008. Epub 2015 Nov 10.
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.
人类非洲锥虫病（HAT）是一种由动基体感染引起的疾病。血清学检测对流行病学监测很有用。本研究的目的是开发一种用于HAT的多重血清学检测方法，以评估所选HAT抗原在血清流行病学监测中的诊断价值。我们克隆了来自布氏冈比亚锥虫的八个抗原的编码基因座，在细菌系统中表达这些基因，并纯化产生的蛋白质。使用来自HAT和VL患者的血清对这些抗原进行Luminex多重检测，以评估这些抗原的免疫诊断潜力。在布氏冈比亚锥虫抗原中，64 kDa和65 kDa不变表面糖蛋白（ISG）以及鞭毛钙结合蛋白（FCaBP）对布氏冈比亚锥虫患者的血清具有高敏感性，在受试者工作特征（ROC）分析中，其曲线下面积（AUC）值分别为0.871、0.737和0.858。ISG64、ISG65和FCaBP抗原与罗德西亚锥虫患者的血清有部分交叉反应。GM6抗原与罗德西亚锥虫患者的血清以及VL患者的血清有交叉反应。此外，还观察到对每种HAT抗原的异质性抗体反应。在同一检测板中检测多种HAT抗原可实现特异性和灵敏的检测。我们的结果证明了应用多重检测方法开发和评估用于血清流行病学监测的HAT抗原的实用性。
