OBJECTIVE: To examine the usefulness of the second lumbrical-interosseous (2L-INT) distal motor latency (DML) comparison test in localizing median neuropathy to the wrist in patients with absent median sensory and motor response in routine nerve conduction studies. METHODS: Electrodiagnostic results from 1,705 hands of patients with carpal tunnel syndrome (CTS) symptoms were reviewed retrospectively. All subjects were evaluated using routine nerve conduction studies: median sensory conduction recorded from digits 1 to 4, motor conduction from the abductor pollicis brevis muscle, and the 2L-INT DML comparison test. RESULTS: Four hundred and one hands from a total of 1,705 were classified as having severe CTS. Among the severe CTS group, 56 hands (14.0%) showed absent median sensory and motor response in a routine nerve conduction study, and, of those hands, 42 (75.0%) showed an abnormal 2L-INT response. CONCLUSION: The 2L-INT DML comparison test proved to be a valuable electrodiagnostic technique in localizing median mononeuropathy at the wrist, even in the most severe CTS patients.