title: Prenatal diagnosis of beta-thalassaemia using fetal erythroblasts enriched from maternal blood by a novel gradient.
abstract: We have assessed a new technique for the isolation of fetal erythroblasts from maternal blood for the non-invasive prenatal diagnosis of pregnancies at risk of beta-thalassaemia. This method relies on the separation of erythroblasts from maternal nucleated cells by a novel step gradient and high speed centrifugation. In four of the six cases examined, single erythroblasts were identified by immunohistochemistry for zeta (zeta) globin. These were individually micromanipulated and analysed by single cell polymerase chain reaction (PCR) and subsequent sequencing of the region of beta-globin locus where the mutations most common to the region of Puglia, Italy, are clustered. In each of the four instances where fetal erythroblasts were identified by antibody staining, the fetal beta-globin genotype was correctly determined. To date, this represents the largest series of non-invasive prenatal diagnoses performed for this haemoglobinopathy.
authors.affiliation: Department of Obstetrics and Gynecology, University of Bari, Italy. edoardodinaro@hotmail.com
authors.forename: E
authors.initials: E
authors.lastname: Di Naro
authors.forename: F
authors.initials: F
authors.lastname: Ghezzi
authors.forename: A
authors.initials: A
authors.lastname: Vitucci
authors.forename: N
authors.initials: N
authors.lastname: Tannoia
authors.forename: D
authors.initials: D
authors.lastname: Campanale
authors.forename: V
authors.initials: V
authors.lastname: D'Addario
authors.forename: W
authors.initials: W
authors.lastname: Holzgreve
authors.forename: S
authors.initials: S
authors.lastname: Hahn
mesh_terms: D002499:Centrifugation, Density Gradient / Q000379:methods*; D015193:Chorionic Villi Sampling; D004900:Erythroblasts / Q000502:physiology*; D005260:Female; D005312:Fetal Blood; D006801:Humans; D008297:Male; D017354:Point Mutation; D016133:Polymerase Chain Reaction; D011247:Pregnancy; D011296:Prenatal Diagnosis / Q000379:methods*; D017086:beta-Thalassemia / Q000175:diagnosis* / Q000235:genetics