A simple method for diagnosing xeroderma pigmentosum variant.

Patients with xeroderma pigmentosum variant have been diagnosed based on a post-replication repair assay using their cells together with their clinical manifestations. We present here an alternative simple method for the diagnosis of xeroderma pigmentosum variant that measures three cellular markers for DNA repair by autoradiography, unscheduled DNA synthesis, recovery of RNA synthesis, and recovery of replicative DNA synthesis after ultraviolet irradiation. Fibroblasts from a patient are plated on three coverslips parallel with normal cells (control). Unscheduled DNA synthesis, recovery of RNA synthesis, and recovery of replicative DNA synthesis after ultraviolet irradiation in the patient's cells are compared with those of adjacent normal cells by counting numbers of grains on nuclei for each coverslip. Of the hereditary photosensitive disorders including xeroderma pigmentosum, Cockayne syndrome, and newly established ultraviolet-sensitive syndrome, only xeroderma pigmentosum variant cells exhibit normal unscheduled DNA synthesis, normal recovery of RNA synthesis, but reduced recovery of replicative DNA synthesis (approximately 50% of that of control cells). This reduction of DNA synthesis is enhanced in the presence of caffeine. Because each disorder yields a different combination of these three markers, this method also provides a systematic basis for the diagnosis of these diseases.