Objective To evaluate the possibility of detection mutations of 〖JP2〗multiple genes in stool for secondary screening for colorectal cancer. Methods Tumor specimens and stool samples from 40 patients with colorectal cancer and 40 normal persons were examined for mutations of p53, K-ras and APC gene by polymerase chain reaction single strand conformation polymorphism (PCR-SSCP) and silver nitrate staining. Results ①The mutation rate of p53, K-ras and APC gene in the tissues and stools of colorectal cancer respectively were 57.50%, 50.00%, 60.00% and 42.86%, 40.00%, 51.43%, and no mutations were found in normal mucosa and stool. ②The mutation ratioes between multiple gene and single gene had significant difference (P＜0.05). ③The sensitivities had no significant difference between faecal occult blood test (FOBT) and multiple gene mutations detection in the diagnosis of colorectal cancer, but the specificity of the latter was higher than FOBT (P＜0.05). Conclusion Detection of multiple gene mutations in stool is a vauble method in the secondary screening for colorectal cancer.