The purpose of this study was to determine the usefulness of salivary biochemical markers for the screening of periodontal disease and examine the agreement between the results of saliva enzyme tests and those of probing depth. The present study included a total of 187 subjects who underwent annual medical check-ups at the Comprehensive Health Care Center, Honjo, Saitama Prefecture, Japan. Periodontal pocket probing was performed with a WHO probe, and various enzymes and biochemical parameters in saliva were measured. For lactate dehydrogenase (LDH), the proportions of the five isoenzymes were calculated. To decide the cut-off point for each enzymatic activity, receiver operating characteristic curves (ROC curves) were constructed and the points of minimum difference between sensitivity and specificity were decided. Among the biochemical markers tested, salivary LDH level had the highest sensitivity and specificity (sensitivity 0.66, specificity 0.67), while salivary levels of aspartate aminotransferase (AST) and blood urea nitrogen (BUN) also had sensitivity and specificity above 0.60. Among the LDH isoenzymes, LDH4 and LDH5 dominated in whole saliva samples. Salivary LDH may be a feasible and useful parameter for the screening of periodontal disease, while salivary AST and BUN also appear to be potentially useful for this purpose.