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Allele-specific methylated multiplex real-time quantitative PCR (ASMM RTQ-PCR), a powerful method for diagnosing loss of imprinting of the 11p15 region in Russell Silver and Beckwith Wiedemann syndromes

Authors

  • Salah Azzi,

    1. APHP, Hôpital Armand Trousseau, Laboratoire d'Explorations Fonctionnelles Endocriniennes; INSERM UMR-S938 Team 4; Université Pierre et Marie Curie-Paris 6, Paris, France
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  • Virginie Steunou,

    1. APHP, Hôpital Armand Trousseau, Laboratoire d'Explorations Fonctionnelles Endocriniennes; INSERM UMR-S938 Team 4; Université Pierre et Marie Curie-Paris 6, Paris, France
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  • Alexandra Rousseau,

    1. Hôpital Saint-Antoine-URCEST, service de Pharmacologie
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  • Sylvie Rossignol,

    1. APHP, Hôpital Armand Trousseau, Laboratoire d'Explorations Fonctionnelles Endocriniennes; INSERM UMR-S938 Team 4; Université Pierre et Marie Curie-Paris 6, Paris, France
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  • Nathalie Thibaud,

    1. APHP, Hôpital Armand Trousseau, Laboratoire d'Explorations Fonctionnelles Endocriniennes; INSERM UMR-S938 Team 4; Université Pierre et Marie Curie-Paris 6, Paris, France
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  • Fabienne Danton,

    1. APHP, Hôpital Armand Trousseau, Laboratoire d'Explorations Fonctionnelles Endocriniennes; INSERM UMR-S938 Team 4; Université Pierre et Marie Curie-Paris 6, Paris, France
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  • Marilyne Le Jule,

    1. APHP, Hôpital Armand Trousseau, Laboratoire d'Explorations Fonctionnelles Endocriniennes; INSERM UMR-S938 Team 4; Université Pierre et Marie Curie-Paris 6, Paris, France
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  • Christine Gicquel,

    1. Epigenetics in Human Health and Disease, Baker IDI Heart and Diabetes Institute, Melbourne, Australia
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  • Yves Le Bouc,

    1. APHP, Hôpital Armand Trousseau, Laboratoire d'Explorations Fonctionnelles Endocriniennes; INSERM UMR-S938 Team 4; Université Pierre et Marie Curie-Paris 6, Paris, France
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  • Irène Netchine

    Corresponding author
    1. APHP, Hôpital Armand Trousseau, Laboratoire d'Explorations Fonctionnelles Endocriniennes; INSERM UMR-S938 Team 4; Université Pierre et Marie Curie-Paris 6, Paris, France
    • Explorations Fonctionnelles Endocriniennes, Hôpital Armand Trousseau, Pierre & Marie Curie School of Medicine, INSERM UMR-S938, 26 Av du Dr Arnold Netter, 75012, France
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  • Communicated by Arupa Ganguly

Abstract

Many human syndromes involve a loss of imprinting (LOI) due to a loss (LOM) or a gain of DNA methylation (GOM). Most LOI occur as mosaics and can therefore be difficult to detect with conventional methods. The human imprinted 11p15 region is crucial for the control of fetal growth, and LOI at this locus is associated with two clinical disorders with opposite phenotypes: Beckwith-Wiedemann syndrome (BWS), characterized by fetal overgrowth and a high risk of tumors, and Russell-Silver syndrome (RSS), characterized by intrauterine and postnatal growth restriction. Until recently, we have been using Southern blotting for the diagnosis of RSS and BWS. We describe here a powerful quantitative technique, allele-specific methylated multiplex real-time quantitative PCR (ASMM RTQ-PCR), for the diagnosis of these two complex disorders. We first checked the specificity of the probes and primers used for ASMM RTQ-PCR. We then carried out statistical validation for this method, on both retrospective and prospective populations of patients. This analysis demonstrated that ASMM RTQ-PCR is more sensitive than Southern blotting for detecting low degree of LOI. Moreover, ASMM RTQ-PCR is a very rapid, reliable, simple, safe, and cost effective method. Hum Mutat 32:249–258, 2011. © 2011 Wiley-Liss, Inc.

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