A MULTIPLE ANTIGEN-DETECTION DIPSTICK COLLOIDAL DYE IMMUNOASSAY FOR THE FIELD DIAGNOSIS OF TRYPANOSOME INFECTIONS IN CATTLE

Monoclonal antibodies (McAbs) were developed against aspartate aminotransferase purified from Trypanosoma brucei rhodesiense bloodstream form (bf) soluble extracts using a combination of anion-exchange and hydrophobic interaction chromatography. McAb 1A1 was Trypanozoon and Nannomonas specific while 2F1 was Trypanozoon bloodstream form specific. A dipstick colloidal dye immunoassay (DIA) was employed as a field diagnostic test for African trypanosome infections and designed using affinity purified polyclonal antibodies (PcAbs) raised against T. b. rhodesiense bf and the two McAbs, 1A1 and 2F1. PcAbs were adsorbed onto Palanil Red dye particles and used as dye reagents. Dipsticks were dotted with the three different antibodies, which captured trypanosomal antigens in samples tested, while the dye reagent bound to the captured antigens; the presence of coloured dots on the dipstick identified trypanosome infections. A field trial of the DIA was carried out in southeastern Uganda. A total of 1686 cattle from seven areas were bled and tested by DIA and haematocrit centrifuge technique (HCT). A total of 798 cattle (47.3%) were found to be trypanosomal antigen positive by DIA while only 162 (9.6%) were revealed to harbour trypanosomes by HCT, of which 151 (93%) were also positive by DIA.