1. Development of a Nanobody-based lateral flow assay to detect active Trypanosoma congolense infections
- Author
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Shaohong Lu, Didier Vertommen, Stefan Magez, Yann G.-J. Sterckx, Rui Chen, Peter Naniima, Serge Muyldermans, Zeng Li, Joar Esteban Pinto Torres, Ding Jianzu, Julie Goossens, Faculty of Sciences and Bioengineering Sciences, Department of Bio-engineering Sciences, and Cellular and Molecular Immunology
- Subjects
0301 basic medicine ,Trypanosoma congolense ,Science ,ANTIGEN ,Pyruvate Kinase ,CATTLE ,Protozoan Proteins ,Heterologous ,Antigens, Protozoan ,GLYCOLYTIC-ENZYMES ,DIAGNOSIS ,Parasitemia ,Extracellular vesicles ,Sensitivity and Specificity ,03 medical and health sciences ,Drug treatment ,Mice ,Antigen ,NORTHERN TOGO ,parasitic diseases ,medicine ,Journal Article ,Medicine and Health Sciences ,Animals ,MEDIATED ISOTHERMAL AMPLIFICATION ,SINGLE-DOMAIN ANTIBODIES ,Immunoassay ,Multidisciplinary ,biology ,Plasma samples ,EXTRACELLULAR VESICLES ,Diagnostic test ,Biology and Life Sciences ,Single-Domain Antibodies ,biology.organism_classification ,medicine.disease ,Virology ,ANIMAL AFRICAN TRYPANOSOMOSIS ,030104 developmental biology ,Trypanosomiasis, African ,general ,Trypanosoma ,Medicine ,Cattle ,Trypanosomiasis ,MOLECULAR SURVEY - Abstract
Animal African trypanosomosis (AAT), a disease affecting livestock, is caused by parasites of the Trypanosoma genus (mainly T. vivax and T. congolense). AAT is widespread in Sub-Saharan Africa, where it continues to impose a heavy socio-economic burden as it renders development of sustainable livestock rearing very strenuous. Active case-finding and the identification of infected animals prior to initiation of drug treatment requires the availability of sensitive and specific diagnostic tests. In this paper, we describe the development of two heterologous sandwich assay formats (ELISA and LFA) for T. congolense detection through the use of Nanobodies (Nbs). The immunisation of an alpaca with a secretome mix from two T. congolense strains resulted in the identification of a Nb pair (Nb44/Nb42) that specifically targets the glycolytic enzyme pyruvate kinase. We demonstrate that the Nb44/Nb42 ELISA and LFA can be employed to detect parasitaemia in plasma samples from experimentally infected mice and cattle and, additionally, that they can serve as ‘test-of-cure’ tools. Altogether, the findings in this paper present the development and evaluation of the first Nb-based antigen detection LFA to identify active T. congolense infections.
- Published
- 2018