Screening test for phenylketonuria
Phenylketonuria (PKU) is a genetic disorder that is characterized by an inabilityof the body to utilize the essential amino acid, phenylalanine. PKU is due to arecessively inherited defect in which the body cannot use the amino acid phenylalanine properly. Phenylalanine, which comes from all dietary protein, accumulates in the blood.The practice of newborn screening was originally developed to detect PKU. Thereare a variety of laboratory tests used in screening for PKU. Elevated levels of phenylalanine are most often detected using a bacterial inhibition (Guthrie) assay, asemiquantitative test.Guthrie test
Method:1.A drop of blood is usually obtained by pricking the heel of a newborninfant in ahospital nurseryon the second or third day of life.2.The blood is collected on a piece of filter paper and mailed to a centrallaboratory.3.A small disk of the filter paper is punched out and placed on anagar gel plate containingBacillus subtilisand B-2-thienylalanine. Each gel holds60-80 disks. The agar gel is able to support bacterial growth but the B-2-thienylalanine inhibits bacterial growth. However, in the presence of extra phenylalanine leached from the impregnated filter paper disk, theinhibition is overcome and the bacteria grow.4.Within a day the bacterial growth surrounding the paper disk is visible tothe eye.5.The amount of growth, measured as the diameter of the colony, is roughly proportional to the amount of phenylalanine in the serum.6.The result is read by comparing the diameter of each sample disk's colonyto the colonies of a series of reference disks with standard phenylalaninecontent included on each large plate.
Interpretation of results1.The Guthrie assay is sensitive enough to detect serum phenylalanine levelsof 180-240 μmol/L (3-4 mg/dL).2.In healthy normal people, phenylalanine levels are usually under 120μmol/L.3.When an elevated level of phenylalanine is detected, the laboratorynotifies the infant's physician, who explains the result to the family andarranges testing to determine the cause of the high phenylalanine levels.