title: Preimplantation diagnosis of alpha-thalassemia by blastomere aspiration and polymerase chain reaction: preliminary experience.
abstract: The ability to diagnose and detect the presence of debilitating and life-threatening diseases would be an invaluable asset to those involved in in vitro fertilization programs. To investigate the possibility of diagnosing severe alpha-thalassemia before implantation, DNA sequences targeting the abnormal deletion breakpoint and the normal region of the alpha-globin gene were amplified by polymerase chain reaction from blastomeres aspirated from human preimplantation embryos. Before performing this experiment, a mouse model blastomere aspiration technique was examined to confirm embryo survival and normal development capability. The mouse embryos showed undisturbed development in the five- to seven-, as well as the eight-cell stages when one, or two (eight-cell stage only) blastomeres were aspirated. The four-cell embryos had a lower survival rate but had a blastocyst formation rate comparable to the more advanced stages and the control embryos. In the human embryo experiment, no homozygous Southeast Asian alpha-thalassemia was detected in the aspirated blastomeres, although two heterozygotic alpha-thalassemia embryos were discovered. The sensitivities of targeting sequence detection in the mouse and human blastomeres were between 50% and 76% in the respective groups. This study supports the feasibility of preimplantation diagnosis of severe alpha-thalassemia when both father and mother are carriers of this inheritable disease.
authors.affiliation: Section of Reproductive Medicine, Department of Obstetrics and Gynecology, Chang Gung Memorial Hospital, Taipei.
authors.forename: M Y
authors.initials: MY
authors.lastname: Chang
authors.forename: Y K
authors.initials: YK
authors.lastname: Soong
authors.forename: M L
authors.initials: ML
authors.lastname: Wang
mesh_terms: D000818:Animals; D001483:Base Sequence; D001757:Blastomeres / Q000502:physiology*; D002681:China / Q000208:ethnology; D005260:Female; D005307:Fertilization in Vitro; D005315:Fetal Diseases; D006801:Humans; D051379:Mice; D008813:Mice, Inbred ICR; D008969:Molecular Sequence Data; D016133:Polymerase Chain Reaction; D011247:Pregnancy; D011296:Prenatal Diagnosis; D012680:Sensitivity and Specificity; D013624:Taiwan; D017085:alpha-Thalassemia