title: Multiple displacement amplification improves PGD for fragile X syndrome.
abstract: We report an improvement in the PGD test for fragile X syndrome (FXS). Recently, multiple displacement amplification (MDA) has been reported to yield large amounts of DNA from single cells. Taking into account this technique, we developed a new PGD test for FXS, enabling combined analysis of linked polymorphic markers with the study of the non-expanded CGG repeat. Single cell amplification efficiency was first assessed on single lymphocytes. Amplification rate of the different markers ranged from 85 to 95% with an allele drop-out (ADO) rate comprised between 7 and 34%. Using this test, eight PGD cycles were carried out for six couples, and 37 embryos were analysed after preliminary MDA. Amplification rate was increased by this technique from 41 to 66% so that embryos with no results were rarer (14 versus 45% without MDA). Reliability of the test was considerably improved by combining direct with indirect genetic analysis. Furthermore, in cases of fully expanded alleles too large to be amplified by PCR, this test gives an internal amplification control. Embryonic transfers were carried out in all but one PGD cycles. One biochemical and one clinical pregnancy resulted, and a healthy child was born. This single diagnosis procedure could be suitable to most patients carrying FXS.
authors.affiliation: Faculté de Médecine, Université Paris-Descartes, Unité INSERM U781 Institut de Recherche Necker-Enfants Malades, Service de génétique médicale, Hôpital Necker-Enfants Malades (Assistance Publique-Hôpitaux de Paris), Paris, France. burlet@necker.fr
authors.forename: P
authors.initials: P
authors.lastname: Burlet
authors.forename: N
authors.initials: N
authors.lastname: Frydman
authors.forename: N
authors.initials: N
authors.lastname: Gigarel
authors.forename: V
authors.initials: V
authors.lastname: Kerbrat
authors.forename: G
authors.initials: G
authors.lastname: Tachdjian
authors.forename: E
authors.initials: E
authors.lastname: Feyereisen
authors.forename: J-P
authors.initials: JP
authors.lastname: Bonnefont
authors.forename: R
authors.initials: R
authors.lastname: Frydman
authors.forename: A
authors.initials: A
authors.lastname: Munnich
authors.forename: J
authors.initials: J
authors.lastname: Steffann
mesh_terms: D004252:DNA Mutational Analysis; D005260:Female; D051860:Fragile X Messenger Ribonucleoprotein 1 / Q000235:genetics; D005600:Fragile X Syndrome / Q000097:blood / Q000175:diagnosis* / Q000235:genetics; D006801:Humans; D016133:Polymerase Chain Reaction / Q000379:methods; D011247:Pregnancy; D019836:Preimplantation Diagnosis / Q000379:methods*; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity
chemical_list: D051860:Fragile X Messenger Ribonucleoprotein 1; C497683:FMR1 protein, human