PURPOSE: A number of pitfalls in single-cell DNA analysis, including undetected DNA contamination, undetected allele drop out, and preferential amplification, may lead to misdiagnosis in preimplantation genetic diagnosis of single-gene disorders. METHODS: Preimplantation genetic diagnosis was performed by sequential first and second polar body analysis of oocytes in 26 couples at risk for having children with various single-gene disorders. Mutant genes were amplified simultaneously with linked polymorphic markers, and only embryos resulting from the mutation-free oocytes predicted by polar body analysis with confirmation by polymorphic marker testing were transferred back to patients. RESULTS: Overall 529 oocytes from 48 clinical cycles (26 patients) were tested, resulting in the transfer of 106 embryos in 44 clinical cycles. As many as 46 (9.6%) instances of allele dropout were observed, the majority (96%) of which were detected. Seventeen unaffected pregnancies were established, of which nine resulted in the birth of an unaffected child, and the rest are ongoing. CONCLUSIONS: A high accuracy of preimplantation genetic diagnosis of single-gene disorders is achieved by application of sequential analysis of the first and second polar body and multiplex polymerase chain reaction.
PURPOSE: A number of pitfalls in single-cell DNA analysis, including undetected DNA contamination, undetected allele drop out, and preferential amplification, may lead to misdiagnosis in preimplantation genetic diagnosis of single-gene disorders. METHODS: Preimplantation genetic diagnosis was performed by sequential first and second polar body analysis of oocytes in 26 couples at risk for having children with various single-gene disorders. Mutant genes were amplified simultaneously with linked polymorphic markers, and only embryos resulting from the mutation-free oocytes predicted by polar body analysis with confirmation by polymorphic marker testing were transferred back to patients. RESULTS: Overall 529 oocytes from 48 clinical cycles (26 patients) were tested, resulting in the transfer of 106 embryos in 44 clinical cycles. As many as 46 (9.6%) instances of allele dropout were observed, the majority (96%) of which were detected. Seventeen unaffected pregnancies were established, of which nine resulted in the birth of an unaffected child, and the rest are ongoing. CONCLUSIONS: A high accuracy of preimplantation genetic diagnosis of single-gene disorders is achieved by application of sequential analysis of the first and second polar body and multiplex polymerase chain reaction.
Authors: A Kuliev; S Rechitsky; O Verlinsky; V Ivakhnenko; J Cieslak; S Evsikov; G Wolf; M Angastiniotis; G Kalakoutis; C Strom; Y Verlinsky Journal: J Assist Reprod Genet Date: 1999-04 Impact factor: 3.412
Authors: Y Verlinsky; S Rechitsky; J Cieslak; V Ivakhnenko; G Wolf; A Lifchez; B Kaplan; J Moise; J Walle; M White; N Ginsberg; C Strom; A Kuliev Journal: Biochem Mol Med Date: 1997-12
Authors: S Rechitsky; C Strom; O Verlinsky; T Amet; V Ivakhnenko; V Kukharenko; A Kuliev; Y Verlinsky Journal: J Assist Reprod Genet Date: 1998-05 Impact factor: 3.412
Authors: A Kuliev; S Rechitsky; O Verlinsky; V Ivakhnenko; S Evsikov; G Wolf; M Angastiniotis; D Georghiou; V Kukharenko; C Strom; Y Verlinsky Journal: J Assist Reprod Genet Date: 1998-05 Impact factor: 3.412