A rapid, simple, and sensitive immunochromatographic test strip has been developed for testing residues of ciprofloxacin (CIP). A specific and sensitive monoclonal antibody (mAb) for CIP was generated by immunizing BALB/c mice with well-characterized CIP-Keyhole limpet haemocyanin. Under the optimized conditions, the cut-off limits of test strips for CIP were found to be 5 ng/mL in phosphate-buffered saline and 2.5 ng/mL in milk samples. Each test can be evaluated within 3 min. The cross-reactivities of the CIP test strip to enrofloxacin (ENR), norfloxacin (NOR), nadifloxacin (NDF), danofloxacin (DANO), pefloxacin (PEX), lomefloxacin (LOME), enoxacin (ENO), and sarafloxacin (SAR) were 71.4%, 71.4%, 66%, 50%, 33%, 20%, 12.5%, and 6.25%, respectively. The data indicate that the method is sensitive, specific, and has the advantages of simplicity and speed, therefore, this test strip is a useful screening method for the detection of CIP residues in milk samples.