Rapid Tests
The Ingezim® ASFV CROM Ag (Eurofins Technologies Ingenasa) is a double antibody sandwich immunochromatographic assay for the detection of ASFV antigen in blood samples. The test is based on the use of two different colored latex microspheres: black microspheres that are coated with a specific monoclonal antibody (MAb) against ASFV p72, and blue microspheres, which are used as test control. On the membrane, two lines are printed: the test line (T) has a specific MAb against ASFV and the control line (C) has a specific MAb against the control protein. In case of a positive sample, the virus binds to the black beads conjugated with the anti-ASFV MAb. The immune complex then migrates through the membrane by capillarity and is captured again by the same anti-ASFV MAb absorbed on the test line, resulting in the appearance of a black line. The presence of the control line serves as validation of the test, indicating that the immunochromatography has been performed correctly (Sastre et al., 2016). The test procedure was conducted according to the manufacturer’s instructions with the exception of also including previously freeze-thawed samples in the study (see supporting tables 1 and 2), using 20 μl of blood, adding 3-4 drops of running buffer after one minute, and results were recorded after ten minutes. The outcome was interpreted either as positive or negative (see figure 1). Only valid results were counted (appearance of the control line).