We reported that endogenous urinary 3-hydroxyproline (3-Hyp) is useful for cancer screening because cancer invasion involves the destruction of basement membrane. A simple and sensitive assay is desired.An ELISA method using a specific antibody against a synthetic peptide of 10 amino acids including 3-Hyp corresponding to the amino acid sequences of collagen type IV alpha chain was applied to urine samples from 180 healthy controls and 22 cancer patients.The values in controls were 2.44+/-1.90 (SD) mg peptide/g creatinine for 52 men and 2.87+/-2.01 for 128 women, while the values in 22 cancer patients were very low at 0.110+/-0.137 (p0.001).The discrepancy in the data between our previous and present studies is based on the difference of targets measured. 3-Hyp-containing peptides in cancer patients might be destroyed by the elevated peptidase levels found in these patients.This ELISA assay may be useful for cancer screening.