title: Multiple displacement amplification for preimplantation genetic diagnosis of  fragile X syndrome.
abstract: Preimplantation genetic diagnosis (PGD) has become an  assisted reproductive technique for couples that have genetic risks. Despite  the many advantages provided by PGD, there are several problems, including  amplification failure, allele drop-out and amplification inefficiency. We  evaluated multiple displacement amplification (MDA) for PGD of the fragile X  syndrome. Whole genome amplification was performed using MDA. MDA products were  subjected to fluorescent PCR of fragile X mental retardation-1 (FMR1)  CGG repeats, amelogenin and two polymorphic markers. In the pre-clinical  tests, the amplification rates of the FMR1 CGG repeat, DXS1215 and  FRAXAC1 were 84.2, 87.5 and 75.0%, respectively, while the allele dropout rates  were 31.3, 57.1 and 50.0%, respectively. In two PGD treatment cycles, 20  embryos among 30 embryos were successfully diagnosed as 10 normal embryos, four  mutated embryos and six heterozygous carriers. Three healthy embryos were  transferred to the uterus; however, no clinical pregnancy was achieved. Our  data indicate that MDA and fluorescent PCR with four loci can be successfully  applied to PGD for fragile X syndrome. Advanced methods for amplification of  minuscule amounts of DNA could improve the sensitivity and reliability of PGD  for complicated single gene disorders.
authors.affiliation: Laboratory of Reproductive Biology and Infertility, Cheil General Hospital & Women's Healthcare Center, Kwandong University College of Medicine, Seoul, Korea. hslee999@gmail.com
authors.forename: H-S
authors.initials: HS
authors.lastname: Lee
authors.forename: M J
authors.initials: MJ
authors.lastname: Kim
authors.forename: C K
authors.initials: CK
authors.lastname: Lim
authors.forename: J W
authors.initials: JW
authors.lastname: Cho
authors.forename: I O
authors.initials: IO
authors.lastname: Song
authors.forename: I S
authors.initials: IS
authors.lastname: Kang
mesh_terms: D000328:Adult; D005260:Female; D051860:Fragile X Messenger Ribonucleoprotein 1 / Q000235:genetics*; D005600:Fragile X Syndrome / Q000175:diagnosis* / Q000235:genetics*; D006801:Humans; D008297:Male; D021141:Nucleic Acid Amplification Techniques / Q000379:methods*; D011247:Pregnancy; D019836:Preimplantation Diagnosis / Q000379:methods*; D018911:Trinucleotide Repeats / Q000235:genetics*
chemical_list: D051860:Fragile X Messenger Ribonucleoprotein 1; C497683:FMR1 protein, human