Preimplantation genetic diagnosis of X-linked adrenoleukodystrophy with gender determination 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 X-linked adrenoleukodystrophy.


METHODS
MDA was used to amplify the whole genome directly from a single blastomere. MDA products were used for polymerase chain reaction (PCR) analysis of two polymorphic markers flanking the ABCD1 gene and a new X/Y marker, X22, to sex embryos in an X-linked adrenoleukodystrophy PGD program.


METHODS
Fertility and gynecology private center in Alicante, Spain.


METHODS
A couple in which the wife is a carrier of the ABCD1 gene mutation (676A-->C) that was previously identified in her family.


METHODS
MDA of single blastomere and PCR tests for PGD.


METHODS
The ability to analyze single blastomeres for X-linked adrenoleukodystrophy using MDA.


RESULTS
The development of an MDA-PGD protocol for X-linked adrenoleukodystrophy allowed for the diagnosis of five embryos. These were biopsied on day 3 of culture and analyzed. One embryo was an affected male and one embryo was a female carrier. Three healthy female embryos were transferred 48 hours after biopsy. Unfortunately, no pregnancy was achieved.


CONCLUSIONS
The MDA technique is useful for overcoming the problem of insufficient genomic DNA in PGD and allows the simultaneous amplification of different targets to perform a diagnosis of any known gene defect and a sexing test by standard methods and conditions.
Lledó
Belén
B
Bernabeu Institute of Fertility and Gynecology, Alicante, Spain. blledo@institutobernabeu.com
Bernabeu
Rafael
R
Ten
Jorge
J
Galán
Francisco M
FM
Cioffi
Luigi
L
D000326:Adrenoleukodystrophy / Q000175:diagnosis* / Q000235:genetics*; D000328:Adult; D005260:Female; D008040:Genetic Linkage / Q000235:genetics; D006801:Humans; D008297:Male; D021141:Nucleic Acid Amplification Techniques / Q000379:methods*; D010375:Pedigree; D016133:Polymerase Chain Reaction / Q000379:methods; D019836:Preimplantation Diagnosis / Q000379:methods*; D019849:Sex Determination Processes