Abstract
Purpose: Fluorescent polymerase chain reaction (PCR) is a multipurpose technique that can be used for diagnosing sex, single-gene defects, and trisomies as well as determining DNA fingerprints from single cells. However, its effectiveness must be assessed before clinical preimplantation genetic diagnosis (PGD) application.
Methods: Single and multiplex fluorescent PCR was applied to single cells and blastomeres.
Results: Fluorescent PCR can be used to diagnose sex from blastomeres and has been successfully applied in a clinical PGD sexing program resulting in a confirmed pregnancy. A further major advantage of fluorescent PCR is the ability to multiplex, providing multiple diagnoses and DNA fingerprints with a high reliability (~75% for trisomy, 86% for DNA fingerprint) and good accuracy (70–80%). Allele dropout in multiplex PCR is ~20% per allele and does not appear to be associated with the fragment size.
Conclusions: Fluorescent PCR is a powerful technique for PGD, and the effects of allele dropout must be considered, particularly in multiplex PCR.
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Findlay, I., Matthews, P. & Quirke, P. Preimplantation Genetic Diagnosis Using Fluorescent Polymerase Chain Reaction: Results and Future Developments. J Assist Reprod Genet 16, 199–206 (1999). https://doi.org/10.1023/A:1020364807226
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DOI: https://doi.org/10.1023/A:1020364807226