One-step multiplex polymerase chain reaction for preimplantation genetic diagnosis of Huntington disease.

OBJECTIVE

To develop a multiplex polymerase chain reaction (PCR) method for Huntington disease (HD) preimplantation genetic diagnosis (PGD) based on the coamplification of CAG repeats and three different polymorphic microsatellites in a single step of PCR.

DESIGN

Techniques and instrumentation.

SETTING

Tertiary clinical and academic medical center.

PATIENT(S): Thirty-six embryos from seven clinical PGD cycles.

INTERVENTION(S): Patients underwent a PGD cycle with transfer of two unaffected embryos on day 5.

MAIN OUTCOME MEASURE(S): PGD based on mutation identification or exclusion testing for at-risk HD carriers.

RESULT(S): Thirty-six embryos from seven clinical PGD cycles were analyzed with the new method here developed, and results were obtained for 34 of them. Two embryos were transferred on day 5, resulting in two singleton pregnancies.

CONCLUSION(S): An interesting application of this approach can be considered for PGD cycles in which numerous markers must be used. We have also used this one-step multiplex method for PGD for other pathological conditions.