Clinical Experience With Preimplantation Genetic Diagnosis of Cystic Fibrosis (Δf508)

Molecular Diagnostics in Preimplantation Genetic Diagnosis

“…The second step involves the diagnostic test itself, which can be performed in a region of the IVF laboratory, an adjacent laboratory equipped to perform molecular analyses or in a completely separate dedicated molecular genetics laboratory equipped to process single cell sam- Various (gender determination to exclude hemizygotes) Nested PCR, agarose gel (ϩ/Ϫ X/Y) X-linked disorders 6 Various (gender determination to exclude hemizygotes) Nested PCR, heteroduplexing Cystic fibrosis 22,38,44,75 Tay-Sachs disease 111 3 bp deletion (⌬F508) 4 bp insertion Nested PCR, allele-specific amplification RhD blood typing 3 Myotonic dystrophy 127 ϩ/Ϫ RhD gene determines Rh status Expansion of (CTG)n trinucleotide repeat Nested PCR, restriction enzyme Cystic fibrosis, 23 Beta thalassemia, 83 Marfan syndrome, 107 Epidermolysis Bullosa, 100 Lesch-Nyhan syndrome, 101 Sickle cell anemia, 102 Fanconi's anemia, 103 Ornithine transcarbamylase deficiency, 104 Spinal muscular atrophy [108][109][110] Various point mutations Deletion. Distinguish between gene and pseudogene Nested PCR, restriction enzyme (2 mutations in 1 fragment)…”

Real-time multiplex analysis of four beta-thalassemia mutations employing surface plasmon resonance and biosensor technology

“…In this case, the speed of the analysis is critical in order to minimize the production of embryos to be successfully tested and implanted. 27 Other advantages of the methodology described in the present paper are: (a) that it is a nonradioactive methodology and (b) that gel electrophoresis and/or dot-spot analysis are not required.…”

Gamete and Embryo Manipulation

“…261 Moreover, it became evident that pregnancy rates after PGD were comparable to those obtained after standard IVF. 262,263 Despite the reassuring clinical results so far, however, the discussion remains as to whether the removal of two cells from a seven-cell-stage embryo (or larger) reduces its capacity to implant more than if only one cell were removed. 264 A recent randomized controlled trial showed that removal of two blastomeres significantly decreases the likelihood of blastocyst formation compared with removal of one blastomere, although this did not translate into a significant difference in implantation rate (23.5% versus 17.3% for one-cell and two-cell biopsy, respectively, P = 0.216).…”