Background : Near-infrared spectroscopy (NIRS) has recognized potential but limited application for non-invasive diagnostic evaluation. Data analysis methodology that reproducibly distinguishes between the presence or absence of physiologic abnormality could broaden clinical application of this optical technique. Methods : Sample data sets from simultaneous NIRS bladder monitoring and invasive urodynamic pressure-flow studies (UDS) are used to illustrate how a diagnostic algorithm is constructed using classification and regression tree (CART) analysis. Misclassification errors of CART and linear discriminant analysis (LDA) are computed and examples of other urological NIRS data likely amenable to CART analysis presented. Results : CART generated a clinically relevant classification algorithm (error 4%) using 46 data sets of changes in chromophore concentration composed of the whole time series without specifying features. LDA did not (error 16%). Using CART NIRS data provided comparable discriminant ability to the UDS diagnostic nomogram for the presence or absence of obstructive pathology (88% specificity, 84% precision). Pilot data examples from children with and without voiding dysfunction and women with mild or severe pelvic floor muscle dysfunction also show potentially diagnostic differences in chromophore concentration. Conclusions : CART analysis can likely be applied in other NIRS monitoring applications intended to classify patients into those with and without pathology.