Diagnostic Techniques employed by MUSCA

1. Card Agglutination Test for Trypanosoma evansi - an antibody-detection assays, is a rapid direct agglutination test which uses formaldehyde-fixed, comassie-stained, freeze dried trypanosomes of T. evansi

2. Blood Smear Examination. This examination uses a drop of blood on a completely degreased slide 1 cm from the edge. The edge of another slide is placed on the first, at an angle of 30-45°. The angled slide is moved along the first with a steady movement drawing the blood evenly on the first slide. The blood is immediately dried in the air, fixed with methyl alcohol and stained. The most common stain used is Giemsa stain.

3. Wet Mount. A drop of blood is placed on a clean, degreased slide, covered with cover glass and immediately examined under the microscope with x20 or x40 dry objective.

4. Microhematocrit Centrifugation Technique. Blood is collected into heparinized capillary tubes (75 x 1.5mm), which is then sealed at the dry end and centrifuged, sealed end down, at 1200rpm for 10 minutes. In MHCT, 2 pieces of glass (25 x 10 x 1.2 mm) are glued to a slide and the capillary tube is placed between them. A cover slip is placed on the top at the level of the buffy coat-plasma interface, where motile trypanosomes are said to be concentrated. The space around the part of the tube is flooded with water and the buffy coat area is examined under the microscope (x100-200)

5. Inoculation into laboratory animals. Blood treated with sodium heparin or EDTA was inoculated intraperitoneally into mice (0.25-0.5ml). Parasitemia in mice was followed every 3 days post inoculation until day 28 by means of wet blood films from blood taken from the tail tip.

6. Polymerase Chain Reaction (PCR).A test performed to evaluate false-negative results to the ELISA


7.Enzyme-Linked Immuno-Sorbent Assay (ELISA) - a sensitive immunoassay that uses an enzyme linked to an antibody as a marker for the detection of a specific protein, especially an antigen or antibody.

NOTES: ELISA and PCR is not part of the routine diagnostic procedure but MUSCA is technically capable of developing the test kit for ELISA given the proper logistics.