Application of a galactosylceramidase microassay method to early prenatal diagnosis of Krabbe's disease.
An enzymatic microassay method was devised to determine galactosylceramidase activities of microsamples prepared from primary cultures of amniotic fluid cells. A piece of freeze-dried cell mass (0.5--1 microgram dry weight) prepared from a colony on the bottom plate of a culture flask, was weighed and incubated in 4.02 microliter of the assay mixture. A small amount of galactose hydrolyzed from galactosylceramide was amplified about 10 000-fold and determined by using an enzymatic amplification reaction, NAD cycling. Microsamples were also prepared from freeze-dried leukocytes and skin fibroblasts. The present method was applied to prenatal diagnosis using amniotic cells as a sample in a case of high-risk pregnancy for Krabbe's disease. The result was confirmed by analysis of fibroblasts and organs from the aborted fetus.
D000653:Amniotic Fluid / Q000166:cytology / Q000201:enzymology*; D002478:Cells, Cultured; D005260:Female; D005347:Fibroblasts / Q000201:enzymology; D005696:Galactosidases / Q000032:analysis*; D005698:Galactosylceramidase / Q000032:analysis*; D006801:Humans; D007231:Infant, Newborn; D007962:Leukocytes / Q000201:enzymology; D007965:Leukodystrophy, Globoid Cell / Q000175:diagnosis* / Q000201:enzymology; D008832:Microchemistry; D011247:Pregnancy; D011296:Prenatal Diagnosis / Q000379:methods*
D005696:Galactosidases; D005698:Galactosylceramidase