title: Prenatal diagnosis of Duchenne muscular dystrophy using a single fetal nucleated erythrocyte in maternal blood.
abstract: We developed a method that allows prenatal diagnosis of Duchenne muscular dystrophy using a single nucleated erythrocyte (NRBC) isolated from maternal blood. Maternal blood was obtained at 8 to 20 weeks of gestation. NRBCs were separated with Percoll using a discontinuous density gradient method and then collected by micromanipulator under microscopic observation. The entire genome of a single cell was amplified by primer extension preamplification (PEP). Sex was determined from a small aliquot of the PEP reaction. After an NRBC was determined to be male and confirmed to be of fetal origin, dystrophin exons 4, 8, 12, 45, 48, 50, and 51 were determined from the same PEP reaction. This diagnostic method using maternal blood is safer than amniocentesis or cordocentesis and can be applied to other X-linked diseases.
authors.affiliation: Department of Obstetrics and Gynecology, Kohnodai Hospital, Japan.
authors.forename: A
authors.initials: A
authors.lastname: Sekizawa
authors.forename: T
authors.initials: T
authors.lastname: Kimura
authors.forename: M
authors.initials: M
authors.lastname: Sasaki
authors.forename: S
authors.initials: S
authors.lastname: Nakamura
authors.forename: R
authors.initials: R
authors.lastname: Kobayashi
authors.forename: T
authors.initials: T
authors.lastname: Sato
mesh_terms: D001483:Base Sequence; D002467:Cell Nucleus / Q000473:pathology; D002469:Cell Separation; D002501:Centrifugation, Zonal; D017931:DNA Primers; D016189:Dystrophin / Q000097:blood / Q000235:genetics*; D004622:Embryo, Mammalian; D004912:Erythrocytes / Q000473:pathology*; D005260:Female; D005333:Fetus; D017353:Gene Deletion; D005865:Gestational Age; D006801:Humans; D008297:Male; D008969:Molecular Sequence Data; D009136:Muscular Dystrophies / Q000097:blood / Q000175:diagnosis* / Q000196:embryology; D016133:Polymerase Chain Reaction / Q000379:methods; D011247:Pregnancy; D011296:Prenatal Diagnosis / Q000379:methods*; D012732:Sex Determination Analysis; D014960:X Chromosome; D014998:Y Chromosome
chemical_list: D017931:DNA Primers; D016189:Dystrophin