This study was conducted in Khartoum North – Sudan to detect antibiotic residues in different poultry tissues (liver, gizzard and muscle) using Bacillus subtilis as a test organism. A total of 213 tissue samples were obtained from a poultry slaughter house used to slaughter poultry from different poultry farms in Khartoum north. The filter paper and whole tissue methods were used and compared using Bacillus subtilis as test organism . The total number of positive samples was 67 (31.4%) The highest percentage of positive samples was detected in the liver tissues 44 (62%) and the least was in the muscles tissue 8 (11.3%). ( ≥ 3mm inhibition zone was regarded positive).There was no significant difference between this standard inhibition zone and the inhibition zone diameter produced by liver samples , but the diameter of the inhibition zone in the gizzard and the muscle vary significantly with the standard inhibition zone set for the test, with p 0≤.05. Kappa statistic was used to measure the agreement between the two method used to detect the antibiotic residues, there was a significant agreement between the two methods with p ≤.05 and the percentage agreement was 78.8%. Also chi square test showed a highly significant association between the two methods. The current study concluded that the poultry meat showed detectable levels of antibiotic residues which may indicate the widespread misuses of antibiotic in poultry farms and the lack of awareness of farmers regarding the recommended withdrawal periods of drugs.