Urinary glycosaminoglycan excretion quantified by an automated semimicro method in specimens conveniently transported...

Current and future treatments for children with mucopolysaccharidosis (MPS) diseases require early, presymptomatic diagnosis, yet existing diagnostic methods to quantitate urinary glycosaminoglycan (GAG) are labor-intensive, and thus not applicable for newborn screening. Direct and rapid quantification of GAG excretion with 1,9-dimethylmethylene blue (DMB) is applicable to small volumes of urine collected, dried, and mailed on a paper matrix (MPS Test). To determine if this assay could be automated, a robotic instrument was programmed to accomplish the procedure; the pilot method simultaneously determined GAG and creatinine concentrations in 10 patient specimens/run. Each analyte is measured in 4 dilutions, thus increasing the operating range to cover a broad spectrum of normal and pathologic levels. Samples and reagents are mixed in a 96-well tray format in approximately 20 min, and densitometric measurements are recorded in less than 60 s. Optical density measurements are electronically transmitted to a desktop computer to select optimal dilutions, identify values above or below the level of reliability, make calculations, and print reports. This automated method was applied to 255 specimens from 101 subjects representing each of the MPS diseases--specifically, types I (n = 126), II (n = 47), III (n = 48), IV (n = 17), VI (n = 14) and VII (n = 3). This method discriminated pathologic elevations of GAG excretion of MPS patients particularly when multiple specimens were available. Patients with non-MPS lysosomal diseases had normal GAG excretion, except for a patient with fucosidosis who had markedly elevated levels. Automation of the direct DMB method provides the key technology necessary for newborn screening for MPS diseases.