title: [Preimplantation genetic diagnosis of Duchenne muscular dystrophy by single cell triplex PCR].
abstract: OBJECTIVE
  To detect two exons of Duchenne muscular dystrophy (DMD) gene and a gender discrimination locus amelogenin gene by single cell triplex PCR, and to evaluate the possibility of this technique for preimplantation genetic diagnosis (PGD) in DMD family with DMD deletion mutation.
  
  
  METHODS
  Single lymphocytes from a normal male, a normal female, two DMD patients (exon 8 and 47 deleted, respectively) and single blastomeres from the couples treated by the in vitro fertilization pre-embryo transfer (IVF-ET) and without family history of DMD were obtained. Exons 8 and 47 of DMD gene were amplified by a triplex PCR assay, the amelogenin gene on X and Y chromosomes were co-amplified to analyze the correlation between embryo gender and deletion status.
  
  
  RESULTS
  In the normal single lymphocytes, the amplification rate of exons 8 and 47 of DMD and amelogenin gene were 93.8%, 93.8%, and 95.3% respectively. The false positive rate was 3.3%. In the exon 8 deleted DMD patient, the amplification rate of exon 47 of DMD and amelogenin gene was 95.8%, and the false positive rate was 3.3%. In the exon 47 deleted DMD patient, the amplification rate of exon 8 of DMD and amelogenin gene was 95.8%, and the false positive rate was 0. In the single blastomeres, the amplification rate of exons 8 and 47 of DMD and amelogenin gene was 82.5%, 80.0% and 77.5%, respectively, and the false positive rate was 0.
  
  
  CONCLUSIONS
  The single cell triplex PCR protocol for the detection of DMD and amelogenin gene is highly sensitive, specific and reliable, and can be used for PGD in those DMD families with DMD deletion mutation.
authors.affiliation: National Laboratory of Medical Genetics, Xiangya Hospital, Central South University, Changsha, China.
authors.forename: Yue-Li
authors.initials: YL
authors.lastname: Wu
authors.forename: Ling-Qian
authors.initials: LQ
authors.lastname: Wu
authors.forename: Yan-Ping
authors.initials: YP
authors.lastname: Li
authors.forename: Dong-E
authors.initials: DE
authors.lastname: Liu
authors.forename: Qiao
authors.initials: Q
authors.lastname: Zeng
authors.forename: Hai-Yan
authors.initials: HY
authors.lastname: Zhu
authors.forename: Qian
authors.initials: Q
authors.lastname: Pan
authors.forename: De-Sheng
authors.initials: DS
authors.lastname: Liang
authors.forename: Hao
authors.initials: H
authors.lastname: Hu
authors.forename: Zhi-Gao
authors.initials: ZG
authors.lastname: Long
authors.forename: Juan
authors.initials: J
authors.lastname: Li
authors.forename: He-Ping
authors.initials: HP
authors.lastname: Dai
authors.forename: Kun
authors.initials: K
authors.lastname: Xia
authors.forename: Jia-Hui
authors.initials: JH
authors.lastname: Xia
mesh_terms: D053523:Amelogenin / Q000235:genetics*; D001757:Blastomeres / Q000166:cytology / Q000378:metabolism; D041321:Chromosomes, Human, X / Q000235:genetics; D041322:Chromosomes, Human, Y / Q000235:genetics; D020732:Cytogenetic Analysis / Q000379:methods; D005091:Exons / Q000235:genetics; D005260:Female; D017353:Gene Deletion; D006801:Humans; D008214:Lymphocytes / Q000166:cytology / Q000378:metabolism; D008297:Male; D020388:Muscular Dystrophy, Duchenne / Q000097:blood / Q000175:diagnosis* / Q000235:genetics; D016133:Polymerase Chain Reaction / Q000379:methods*; D011247:Pregnancy; D019836:Preimplantation Diagnosis / Q000379:methods*
chemical_list: D053523:Amelogenin