The invention provides a hemoglobin, hemoglobin-haptoglobin composite and transferrin joint examination kit. The hemoglobin, hemoglobin-haptoglobin composite and transferrin joint examination kit comprises an upper shell, a lower shell, an immunostrip, test paper and the like; a gold-labeled composite containing a hemoglobin monoclonal antibody, a hemoglobin-haptoglobin composite monoclonal antibody and a transferrin monoclonal antibody is scribed on a nitrocellulose membrane; 3 test lines, a quality control line (15) and a gold-labeled composite membrane scribing line (8) are arranged on the nitrocellulose membrane in parallel. The invention further provides a preparation method and a detection method of the hemoglobin, hemoglobin-haptoglobin composite and transferrin joint examination kit. The hemoglobin, hemoglobin-haptoglobin composite and transferrin joint examination kit is convenient and simple in operation, stable in performance and accurate in result, has a relatively good reference value for early diagnosis and identification of colorectal cancer or colon cancer tumor or other tumors with lower gastrointestinal bleeding symptoms in clinic, significantly improves the positive detection rate of digestive hemorrhagic diseases, is suitable for clinical hospital examination and household self-examination, and provides measures for large-scale general examination of such diseases.