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. Germline mutation c.-65_-55dup11 (mapped to c.-61_-51dup) was identified in prodand 221, 230 and 231. Proband 221 (12Y) presented with retinal capillary hemangioma. Proband 230 (40Y) presented with CNS hemangioblastoma. Proband 231 (9Y) presented with CNS hemangioblastoma. All three patients were previously published in PMID 17006605.