| Reliable gender screening for human preimplantation embryos, using multiple DNA target-sequences. | |
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MedLine Citation:
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PMID: 1479016 Owner: NLM Status: MEDLINE |
Abstract/OtherAbstract:
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Dependable methods were developed for preimplantation sexing of human IVF embryos, for use in clinical settings where prospective parents are at high risk for transmission of X-linked diseases. Using single cultured cells and blastomeres from human embryos as model systems, a multiplex protocol was developed for rapid analysis via nested polymerase chain reaction (PCR). Reliability was enhanced by co-amplification of conserved amelogenin gene segments from both X and Y chromosomes, as well as Y-linked DYZ1 repetitive elements. Each cell was manually isolated and individually washed to avoid potential contaminants. Multiplex amplification allowed recognition of spurious amplification failures specific to particular amelogenin single-copy targets. The X-linked internal control and multiple Y-linked markers allowed recognition and exclusion of most aberrant samples, thus averting potential misdiagnosis. The optimized single-cell protocol reduced experimental sexing errors to < 2% (1/60), but also revealed potential pitfalls of single-cell analysis. With human triploid embryos, separate sampling of individual blastomeres provided concordant female or male signals. Slight modification adapted the procedure for diagnosis of biopsy material from blastocyst stage embryos, allowing separate analysis of multiple tubes containing multiple cells for improved reliability. |
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Authors:
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G Levinson; R A Fields; G L Harton; F T Palmer; A Maddalena; E F Fugger; J D Schulman |
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Publication Detail:
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Type: Journal Article |
Journal Detail:
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Title: Human reproduction (Oxford, England) Volume: 7 ISSN: 0268-1161 ISO Abbreviation: Hum. Reprod. Publication Date: 1992 Oct |
Date Detail:
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Created Date: 1993-02-05 Completed Date: 1993-02-05 Revised Date: 2006-11-15 |
Medline Journal Info:
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Nlm Unique ID: 8701199 Medline TA: Hum Reprod Country: ENGLAND |
Other Details:
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Languages: eng Pagination: 1304-13 Citation Subset: IM |
Affiliation:
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Genetics and IVF Institute, Fairfax, VA 22031. |
Export Citation:
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APA/MLA Format Download EndNote Download BibTex |
| MeSH Terms | |
Descriptor/Qualifier:
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Amelogenin Base Sequence Blastocyst / ultrastructure* Cells, Cultured DNA / analysis*, isolation & purification DNA, Satellite / analysis Dental Enamel Proteins / genetics Electrophoresis, Polyacrylamide Gel Fertilization in Vitro Humans Linkage (Genetics) Molecular Sequence Data Multigene Family Polymerase Chain Reaction Prenatal Diagnosis* Sensitivity and Specificity Sex Determination (Analysis) / methods* |
| Chemical | |
Reg. No./Substance:
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0/Amelogenin; 0/DNA, Satellite; 0/Dental Enamel Proteins; 9007-49-2/DNA |
From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine
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