title: Preimplantation genetic diagnosis for a Chinese family with autosomal recessive Meckel-Gruber syndrome type 3 (MKS3).
abstract: Meckel-Gruber syndrome type 3 is an autosomal recessive genetic defect caused by mutations in TMEM67 gene. In our previous study, we have identified a homozygous TMEM67 mutation in a Chinese family exhibiting clinical characteristics of MKS3, which provided a ground for further PGD procedure. Here we report the development and the first clinical application of the PGD for this MKS3 family. Molecular analysis protocol for clinical PGD procedure was established using 50 single cells in pre-clinical set-up. After whole genomic amplification by multiple displacement amplification with the DNA from single cells, three techniques were applied simultaneously to increase the accuracy and reliability of genetic diagnosis in single blastomere, including real-time PCR with Taq Man-MGB probe, haplotype analysis with polymorphic STR markers and Sanger sequencing. In the clinical PGD cycle, nine embryos at cleavage-stage were biopsied and subjected to genetic diagnosis. Two embryos diagnosed as free of TMEM67 mutation were transferred and one achieving normal pregnancy. Non-invasive prenatal assessment of trisomy 13, 18 and 21 by multiplex DNA sequencing at 18 weeks' gestation excluded the aneuploidy of the analyzed chromosomes. A healthy boy was delivered by cesarean section at 39 weeks' gestation. DNA sequencing from his cord blood confirmed the result of genetic analysis in the PGD cycle. The protocol developed in this study was proved to be rapid and safe for the detection of monogenic mutations in clinical PGD cycle.
authors.affiliation: Department of Obstetrics and Gynecology, Chinese PLA General Hospital, Beijing, China.
authors.forename: Yanping
authors.initials: Y
authors.lastname: Lu
authors.forename: Hongmei
authors.initials: H
authors.lastname: Peng
authors.forename: Zhanguo
authors.initials: Z
authors.lastname: Jin
authors.forename: Jing
authors.initials: J
authors.lastname: Cheng
authors.forename: Shufang
authors.initials: S
authors.lastname: Wang
authors.forename: Minyue
authors.initials: M
authors.lastname: Ma
authors.forename: Yu
authors.initials: Y
authors.lastname: Lu
authors.forename: Dongyi
authors.initials: D
authors.lastname: Han
authors.forename: Yuanqing
authors.initials: Y
authors.lastname: Yao
authors.forename: Yali
authors.initials: Y
authors.lastname: Li
authors.forename: Huijun
authors.initials: H
authors.lastname: Yuan
mesh_terms: D000328:Adult; D002925:Ciliary Motility Disorders / Q000235:genetics*; D004677:Encephalocele / Q000235:genetics*; D005260:Female; D006239:Haplotypes; D006801:Humans; D008297:Male; D008565:Membrane Proteins / Q000235:genetics*; D009154:Mutation; D007690:Polycystic Kidney Diseases / Q000235:genetics*; D011247:Pregnancy; D019836:Preimplantation Diagnosis / Q000379:methods
chemical_list: D008565:Membrane Proteins; C468846:TMEM67 protein, human
references.citation: Taiwan J Obstet Gynecol. 2007 Mar;46(1):9-14
references.pmid: 17389183
references.citation: Hum Reprod. 2011 Nov;26(11):2915-7
references.pmid: 21840907
references.citation: Mol Neurobiol. 2011 Feb;43(1):12-26
references.pmid: 21110233
references.citation: Hum Reprod. 2010 Nov;25(11):2685-707
references.pmid: 20813804
references.citation: Prenat Diagn. 2001 Mar;21(3):223-32
references.pmid: 11260612
references.citation: Hum Mutat. 2002 May;19(5):554-9
references.pmid: 11968088
references.citation: Fertil Steril. 1998 May;69(5):904-8
references.pmid: 9591501
references.citation: J Cell Biol. 2011 Mar 21;192(6):1023-41
references.pmid: 21422230
references.citation: Biotechnology (N Y). 1992 Apr;10(4):413-7
references.pmid: 1368485
references.citation: Reprod Biomed Online. 2010 Jan;20(1):75-82
references.pmid: 20158991
references.citation: Fetal Diagn Ther. 2010;27(1):1-7
references.pmid: 20051662
references.citation: Proc Natl Acad Sci U S A. 2002 Apr 16;99(8):5261-6
references.pmid: 11959976