This study compared the detection of ASFV nucleic acid from pigs with clinically and pathologically compatible signs between four different sample types: diaphragm meat juice, diaphragm muscle swab, spleen, and spleen swabs. Pigs slaughtered at abattoirs around Kampala, Uganda were evaluated by veterinarians for signs of diarrhea, ecchymosis on the skin, enlarged and hemorrhagic spleens and lymph nodes, and kidney petechiation. If they exhibited at least three of these signs at the time of slaughter, they were included in the study. Diagnostic procedures were done using the US Department of Agriculture’s Foreign Animal Disease Diagnostic Laboratory’s Standard Operating Procedures. The Qiagen DNeasy kit was used for nucleic acid extraction and a real-time polymerase chain reaction assay previously described by Zsak et al. (2004) was used for amplification and detection of the target viral nucleic acid. Of the 501 pigs evaluated, diaphragm meat juice samples classified 365 (72.9%) as positive for ASFV, diaphragm muscle swabs classified 221 (44.1%), spleen samples classified 254 (50.7%) and spleen swabs classified 247 (49.3%) as positive. Meat juice had a statistically greater proportion classified as positive than other samples, and muscle swabs had a statistically lesser proportion classified as positive than all other samples. Meat juice samples provide a reliable sample type for the diagnosis of ASF when using real time PCR.