Hyperphenylalaninemia Due to Dihydropteridine Reductase Deficiency: Diagnosis by Enzyme Assays on Dried Blood Spots

Abstract

Enzymatic diagnosis of hyperphenylalaninemia due to a deficiency of dihydropteridine reductase (DHPR) has previously been made by assay on liver biopsy samples, cultured skin fibroblasts, cultured lymphoid cell lines, or peripheral leukocytes. These procedures have some disadvantages for the purpose of early diagnosis of the disease. A simple method of DHPR assay using erythrocytes or dried blood spots on filter papers is described. The mean DHPR activity erythrocytes of control subjects was 3.20 ± 0.70 (SD) nmoles/min/mg of hemoglobin, those of two patients were undetectable, and those of obligate heterozygotes for DHPR deficiency were approximately 50% of the mean control value. The assay on erythrocytes required only a 5-µl volume of whole blood for one test. The DHPR activities in dried blood spots on filter papers from 100 normal newborns were 5.77 ± 1.16 nmoles/min per 5-mm diameter disc; those from normal older infants, children, and adults were 3.37 ± 0.72 nmoles/min per disc; and those from two adolescent patients with DHPR deficiency were undetectable. No false-positive results were obtained. The stability of DHPR in dried blood on filter papers was enough to mail samples in an ordinary form to a specialist laboratory. The DHPR assay on erythrocytes of dried blood spots can be easily applied to all newborn infants with hyperphenylalaninemia detected using the Guthrie tests, and will facilitate the quick and confirmative detection of DHPR deficiency among them.