Abstract
Background:
The development of technologies that detect monogenic diseases in embryonic and fetal samples are opening novel diagnostic possibilities for preimplantation genetic diagnosis (PGD) and prenatal diagnosis (PND) thereby changing laboratory practice. Molecular diagnostic laboratories use different workflows for PND depending on the disease, type of biological sample, the presence of one or more known mutations, and the availability of the proband. Paternity verification and contamination analysis are also performed. The aim of this study was to test the efficacy of a single workflow designed to optimize the molecular diagnosis of monogenic disease in families at-risk of transmitting a genetic alteration.
Methods:
We used this strategy, which we designated “SEeMORE strategy” (Single-tube Electrophoresis analysis-based genotyping to detect MOnogenic diseases Rapidly and Effectively from conception to birth). It consists of a multiplex PCR that simultaneously carries out linkage analysis, direct analysis, maternal contamination and parenthood testing. We analyzed samples from previously diagnosed families for PND (cystic fibrosis or Duchenne muscular dystrophy) without, however, knowing the results.
Results:
The results obtained with the SEeMORE strategy concurred with those obtained with traditional PND. In addition, this strategy has several advantages: (i) use of one or a few cells; (ii) reduction of the procedure to 1 day; and (iii) a reduction of at least 2–3-fold of the analytic cost.
Conclusions:
The SEeMORE strategy is effective for the molecular diagnosis of monogenic diseases, irrespective of the amount of starting material and of the disease mutation, and can be used for PND and PGD.
Acknowledgments
The authors thank Jean Ann Gilder (Scientific Communication srl., Naples, Italy) for editing the text, and Vittorio Lucignano, CEINGE-Biotecnologie Avanzate, for technical assistance.
Author contributions: All the authors have accepted responsibility for the entire content of this submitted manuscript and have approved submission.
Research Funding: This work was supported by POR CAMPANIA FSE 2007–2013 Project DIAINTECH, Italy (to F.S.); Società Italiana di Biochimica Clinica e Biologia Molecolare Clinica (grant 08/14) (to R.T.); and Consorzio Interuniversitario Biotecnologie (grant 11/15) (to F.C.).
Employment or leadership: None declared.
Honorarium: None declared.
Competing interests: The funding organization(s) played no role in the study design; in the collection, analysis, and interpretation of data; in the writing of the report; or in the decision to submit the report for publication.
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