In a study of 426 unrelated VHL patients, 111 were discovered to have alterations in the VHL gene. 18 novel variants were identified in VHL patients, but none were present in 200 unaffected control individuals. DNA samples from peripheral blood leukocytes underwent polymerase chain reaction (PCR) amplification, denaturing high-performance liquid chromatography (DHPLC), and direct sequence analysis of the entire coding sequence and intron-exon boundaries. The V66G (c.197T>G) germline mutation of the VHL gene was found in a 38 yr old VHL patient with CNS hemangioblastoma, pheochromocytoma, renal cystic lesion and pancreatic cyst or tumor (index case 379). Transmission of the variant could not be determined (n/d) since both parents and family history were not available.