Pitfalls in the application of enzyme-linked immunoassays for the detection of circulating trypanosomal antigens in serum samples

Abstract

The experimental infection of two goats with Trypanosoma vivax trypanosomes provided samples for analysis using parasitology techniques and antigen-detection enzyme-linked immunosorbent assays (ELISAs) for T. vivax, T. congolense and T. brucei. Clinical, parasitological and serological findings were monitored during the course of infection to identify problems in the application of these ELISAs. The data clearly showed that the ELISAs examined were entirely unsuitable for the reliable detection of trypanosomal antigen. Consequently, research strategies pertinent to the development of a new generation of both antigen and antibody ELISAs are outlined considering the problems encountered. These were (1) the reactivity of the reagents; (2) the specificity of the reagents; (3) the nature of the test sample, e.g. the compartmentalisation of trypanosomes between plasma, serum and red blood cells; (4) possible interference with the ELISA through immune complexing; and (5) the biology of the host/trypanosome relationship to gain an understanding of fluctuations in trypanosomes in the systemic circulation.