A validation test piece is for validating a bio-sample detection device, which includes an insert port for insertion of the validation test piece and a detection module. The validation test piece includes a base seat, a test element, a standardized data and a top cover. The base seat includes a seat body and a receiving member disposed on the seat body. The test element is connected to the receiving member, and includes a test region to be detected by the detection module. The standardized data corresponds to a sample number of the test region. The top cover is connected to the base seat for covering the test element.