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.


METHODS
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.


METHODS
Fertility and gynecology private center in Alicante, Spain.


METHODS
A couple in which the husband is affected by Marfan syndrome and carries a novel mutation in the FBN-1 gene.


METHODS
The MDA of single cells and PCR tests for PGD.


METHODS
Allele drop-out (ADO), amplification efficiency rates, and the ability to detect Marfan syndrome using MDA.


RESULTS
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.


CONCLUSIONS
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.
Lledó
Belén
B
Bernabeu Institute of Fertility and Gynecology, Alicante, Spain. belen.lledo@ua.es
Ten
Jorge
J
Galán
Francisco M
FM
Bernabeu
Rafael
R
D000328:Adult; D002874:Chromosome Mapping / Q000379:methods*; D005260:Female; D020022:Genetic Predisposition to Disease / Q000235:genetics; D006801:Humans; D008297:Male; D008382:Marfan Syndrome / Q000175:diagnosis* / Q000235:genetics*; D021141:Nucleic Acid Amplification Techniques / Q000379:methods*; D019836:Preimplantation Diagnosis / Q000379:methods*