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Oxytetracycline has been widely used in the cattle industry to control pneumonia, shipping fever, foot rot, bacterial enteritis, and uterine infections. Extensive use of antibiotics in veterinary clinics has resulted in residues in tissue and bacterial resistance to antibiotics. To prevent unwanted drug residues from entering the human food chain, extensive control measures have been established by both government authorities and industries. The demands for reliable, simple, sensitive, rapid and low-cost methods for residue analysis of foods are increasing. In this study, we established a rapid test for tissue residues of oxytetracycline in cattle. The recommended therapeutic dose of oxytetracycline (withdrawal time, 14 days) was administered to 10 cattle. Blood samples were collected from each cow before drug administration and during the withdrawal period. The concentration of oxytetracycline in plasma, determined by a semi-quantitative ELISA, was compared to that of the internal standard, 10 ppb. The absorbance ratio of internal standard to sample (B/Bs) was employed as an index to determine whether the residues in cattle tissues were negative or positive. That is, a B/Bs ratio less than 1 was considered as residue positive and that greater than 1 as negative. Based on this criterion, all plasma samples from cattle were negative to oxytetracycline at pre-treatment. Oxytetracycline could be detected in the plasma treated cattle until day 14 post-treatment. The present study showed that the semi-quantitative ELISA could be easily adapted in predicting tissue residues for oxytetracycline in live cattle.