To develop a novel peptide probe for imaging detection of colorectal cancer, a 12-mer phage display library was used to select peptides that bind specifically to the human colorectal cancer cell line Caco-2. After four rounds of panning, four phage clones that bound specifically to the Caco-2 cells were selected. The phage clone SP-2 had a particularly high affinity and specificity for Caco-2 cells. This clone was identified using a series of methods. The peptide SP-2 that was displayed on phage SP-2 exhibited a high specificity to Caco-2 cells. Thus, the peptide SP-2 could be a candidate probe for the detection of colorectal cancer.