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.


METHODS
Techniques and instrumentation.


METHODS
Tertiary clinical and academic medical center.


METHODS
Thirty-six embryos from seven clinical PGD cycles.


METHODS
Patients underwent a PGD cycle with transfer of two unaffected embryos on day 5.


METHODS
PGD based on mutation identification or exclusion testing for at-risk HD carriers.


RESULTS
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.


CONCLUSIONS
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.
Peciña
Ana
A
Unidad de Gestión Clínica de Genética, Reproducción y Medicina Fetal, Hospitales Universitarios Virgen del Rocío, 41013 Seville, Spain,
Lozano Arana
María Dolores
MD
García-Lozano
Juan C
JC
Borrego
Salud
S
Antiñolo
Guillermo
G
D001755:Blastocyst / Q000166:cytology / Q000378:metabolism; D001757:Blastomeres / Q000166:cytology / Q000378:metabolism; D015193:Chorionic Villi Sampling; D005260:Female; D006801:Humans; D006816:Huntington Disease / Q000175:diagnosis* / Q000235:genetics; D016133:Polymerase Chain Reaction / Q000295:instrumentation / Q000379:methods*; D011110:Polymorphism, Genetic; D011247:Pregnancy; D019836:Preimplantation Diagnosis / Q000295:instrumentation / Q000379:methods*; D018911:Trinucleotide Repeats / Q000235:genetics