Introduction: The ESHRE guidelines of best practice recommends assay validation on single cells from the family prior to the PGD cycle for each new disease and family in order to demonstrate the efficiency of the multiplex single cell PCR assay.
Aims: To compare the efficiency of the multiplex PGD assay in cases for which single cell validation was and was not performed prior to the first cycle.
Materials and Methods: Retrospective study of all PGD assays for new diseases and new families performed in Shaare Zedek PGD Unit since 2005. Efficiency of the multiplex assay was compared between cases with and without prior fibroblast validation.
Results: Single fibroblast validation was performed in 169 different assays for 82 diseases. The mean number of markers used in each single fibroblast validation was 9.5+4. Thirty eight PGD cycles for new diseases were performed without prior assay validation. The mean number of markers used in each cycle was 7.1+2. The total amplification rate, both for fibroblast validation assays and for PGD cycles without prior validation was 97%. Results were possible in 100% of all cycles even when prior validation was not performed.
Conclusions: Amplification rates for multiplex PCR PGD assays were equally efficient in cases performed with and without prior validation in single cells. In light of these results, we suggest that an experienced PGD lab, which can demonstrate the ability to plan and carry out robust multiplex PCR assays, need not validate these assays in patient cells prior to the PGD cycle.
