Objective: To develop and apply efficient and reliable protocols for preimplantation genetic diagnosis (PGD) for Marfan syndrome.
Design: Two mutation-specific protocols were developed, and the markers D15S1028, D15S992, D15S196, D15S576, D15S123, and D15S143 were used to set up four multiplex polymerase chain reactions (PCRs).
Setting: Research Center Reproduction and Genetics.
Patient(s): Ten couples carrying mutations in the FBN1 gene.
Intervention(s): Six PGD protocols were developed for 10 couples, and 7 of them underwent a total of 16 clinical cycles.
Main outcome measure(s): Amplification, allele drop-out (ADO), and contamination rates during the preclinical assays. DNA analyses of blastomeres from embryos biopsied during PGD cycles.
Result(s): Six different protocols were set up, with the main objective being to to use one protocol for several couples. A total of 16 PGD cycles were performed, which resulted in the delivery of an unaffected boy and three ongoing pregnancies.
Conclusion(s): The development of single-cell multiplex PCRs for linked markers and its use in PGD reduce the workload of the genetic diagnostic laboratory as well as the average waiting time for patients. This approach also allows for the simultaneous and accurate detection of recombination, contamination, and ADO, thereby increasing the reliability of the diagnosis.