The invention relates to the technical field of medicine detection, and discloses a detection card of thiamphenicol. The detect card comprises a detection card body, a detection board is arranged in an inner cavity of the detection card body, location holes are formed in two sides of the detection board, a detection window is arranged in the middle of the inner cavity of the detection board, the inner cavity of the detect window is fixedly connected to a test body, a detection tube is fixedly connected to the top of the test body, and the top of the test body is fixedly connected to a flow guide pipeline. According to the detection method and detection card of thiamphenicol, through cooperative use of the detection body, the location holes and the detection board, a function of stabilizing the detection window can be achieved; then a solution is added into the test body through a sample adding hole; through cooperative use of the detection tube and the flow guide pipeline, more convenience is then brought to a user when the user is conducting a detection, and an occurrence of errors in a detection result caused by movement of the detection tube is avoided; meanwhile, the detection tube and the test body present T-shapes, thus flow velocity of a solution sample is slowed down, not only occurrence of detection failure is prevented, but also detection efficiency is improved.