Reliability of detection by polymerase chain reaction of the sickle cell-containing region of the beta-globin gene in single human blastomeres.
Human preimplantation embryos at various stages of development have been analysed using the polymerase chain reaction to amplify a 680 base pair fragment of the beta-globin gene. Successful amplification was achieved more frequently with DNA from intact embryos containing between one and 11 cells, single cumulus cells, oocytes which had failed to fertilize and polar bodies than from single blastomeres disaggregated from intact embryos and treated in an identical manner. The distribution of nuclei demonstrated using the nuclear chromophore diamino-phenyl-indole showed considerable inter-blastomere variation; however, no clear correlation between staining pattern and successful amplification was observed. The reason for the unreliable amplification of DNA from single blastomeres is unclear but this finding has important implications for preimplantation diagnosis of genetic disease.
Pickering
S J
SJ
Department of Anatomy, University of Cambridge, UK.
McConnell
J M
JM
Johnson
M H
MH
Braude
P R
PR
D001757:Blastomeres / Q000502:physiology*; D002874:Chromosome Mapping; D004247:DNA / Q000235:genetics; D015894:Genome, Human; D005914:Globins / Q000235:genetics*; D006801:Humans; D009865:Oocytes / Q000648:ultrastructure; D016133:Polymerase Chain Reaction; D011296:Prenatal Diagnosis / Q000379:methods*; D015203:Reproducibility of Results; D012805:Sickle Cell Trait / Q000175:diagnosis* / Q000235:genetics
D005914:Globins; D004247:DNA