A specific spectrophotometric assay of muscle-pyruvate kinase (M-PK) was used to measure the activity of this isozyme in fetal muscle, fetal plasma and amniotic fluid at about 17-24 wk of gestational age to assess its predictive value for the prenatal diagnosis of Duchenne muscular dystrophy (DMD). Fetal muscle obtained after termination was found to contain a high M-PK specific activity. No significant activity was detected in amniotic fluid from normal or at-risk fetuses. Pure fetal blood was obtained in utero by fetoscopy; significant plasma levels of M-PK activity were measured in a series of control samples, but at-risk fetal plasma contained no higher levels. We conclude that M-PK is of no use for the prenatal diagnosis of DMD.