Document Detail

Archival fixed and analysed preimplantation human embryonic cells: a DNA resource for retrospective PCR analysis at the cellular level.
MedLine Citation:
PMID:  9239719     Owner:  NLM     Status:  MEDLINE    
Biopsies of human embryonic cell preparations previously analysed by cytogenetic and/or fluorescent in-situ hybridization (FISH) chromosome probes provide a unique reference DNA resource for the archival preimplantation genetic diagnosis (PGD) of the transferred embryo. DNA polymerase chain reaction (PCR) may be utilized on these fixed cell preparations to verify equivocal FISH/PGD results. Retrospective PCR screens of the genotype of biopsied embryonic cell(s) may be of benefit in the case of a suspected genetic mutation. Currently, carrier detection or linkage analysis is often not possible because of early death of the fetus, or of patients with a lethal disease. Alternatively, fixed/stained 'failed fertilized' oocytes provide a resource to extend genetic analysis to infertile patients. A successful research is described which minimizes loss of individual analysed fixed/stained oocytes, metaphase chromosomes, and embryonic cell samples. Initial DNA amplification takes place in situ using a modified PCR protocol. Comparative cellular studies using primer sets previously used for PGD analyses show that 65% of the preparations amplified unequivocally using the modified protocol and primers for a CA repeat motif gene sequence, in comparison with 81% using the original PCR protocol. With further refinement and optimization, the methods outlined have the potential to retrospectively screen archival fixed chromosomes, gametes, and embryonic cells for clinical application, and enable the further study of the fixed human preimplantation embryo at the morphological, cell and molecular level.
T A Smith; M B Wall; A L Muggleton-Harris
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Publication Detail:
Type:  Journal Article; Research Support, Non-U.S. Gov't    
Journal Detail:
Title:  Molecular human reproduction     Volume:  3     ISSN:  1360-9947     ISO Abbreviation:  Mol. Hum. Reprod.     Publication Date:  1997 Feb 
Date Detail:
Created Date:  1997-09-19     Completed Date:  1997-09-19     Revised Date:  2006-11-15    
Medline Journal Info:
Nlm Unique ID:  9513710     Medline TA:  Mol Hum Reprod     Country:  ENGLAND    
Other Details:
Languages:  eng     Pagination:  149-55     Citation Subset:  IM    
Division of Obstetrics and Gynecology, UMDS, St Thomas's Hospital, London, UK.
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MeSH Terms
Blastocyst / chemistry*
Cleavage Stage, Ovum / chemistry*
DNA / genetics*,  isolation & purification*
Embryo Transfer
Genetic Diseases, Inborn / diagnosis,  genetics
Oocytes / chemistry
Polymerase Chain Reaction / methods*
Prenatal Diagnosis / methods
Retrospective Studies
Reg. No./Substance:

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