Preimplantation genetic diagnosis of Marfan syndrome using multiple displacement amplification

Objective: To evaluate the use of multiple displacement amplification (MDA) for whole-genome amplification in the preimplantation genetic diagnosis (PGD) of Marfan syndrome. Design: Multiple displacement amplification was used to amplify the whole-genome directly from a single-cell. The MDA product was used for polymerase chain reaction (PCR) analysis of five different loci. At this point MDA was used to develop a PGD-Marfan syndrome program. Setting: Fertility and gynecology private center in Alicante, Spain. Patient(s): A couple in which the husband is affected by Marfan syndrome and carries a novel mutation in the FBN-1 gene. Intervention(s): The MDA of single cells and PCR tests for PGD. Main Outcome Measure(s): Allele drop-out (ADO), amplification efficiency rates, and the ability to detect Marfan syndrome using MDA. Result(s): We report that isothermal whole-genome amplification from single cells allowed analysis of five different loci using standard conditions. The development of a MDA-PGD protocol for Marfan syndrome allowed for the diagnosis of seven embryos. These were biopsied on day 3 of culture and analyzed. Two healthy embryos were transferred 48 hours after culture, resulting in a singleton ongoing pregnancy and the birth of a healthy child. Conclusion(s): The MDA technique is useful for overcoming the problem of insufficient genomic DNA in PGD. The use of MDA as a universal step marks a new cycle for PGD as it allows for the diagnosis of any known gene defect by standard methods and conditions.