Coughing induces cardioacceleratory responses under cholinergic control. The Cough Test (CT), a standardized test that uses a series of coughs with electrocardiographic monitoring, was used to assess the functional integrity of cardiovascular autonomic nerves. In 224 control subjects and 235 diabetic patients, heart-rate (HR) responses were compared with four established tests: lying to standing (LS), standing to lying (SL), deep breathing (DB), and Valsalva maneuver (VM). In control subjects, HR responses declined significantly with age in a curvilinear pattern. Log-transformed indices were used to estimate percentiles. The CT-HR responses were reproducible and significantly associated with other HR-response tests. Sixty-nine (29%) diabetic patients had an abnormal value less than or equal to 1st percentile, whereas only 2 control subjects had such an abnormality. Among diabetic patients, age and duration of diabetes exerted a significant negative influence. With the use of the criteria of two abnormal or one abnormal and two borderline tests (among CT, LS, SL, DB, or VM) as a minimal criteria for cardiovascular autonomic neuropathy, CT and LS had the least overlap between the control and diabetic populations and were significantly better than SL or VM (P less than 0.005 for CT, P less than 0.01 for LS). They were not different from DB. We found the CT to be simple to perform, reproducible, and useful for the assessment of cardiovascular autonomic reflexes.