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Clinical Chemistry and Laboratory Medicine (CCLM)

Published in Association with the European Federation of Clinical Chemistry and Laboratory Medicine (EFLM)

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Ed. by Gillery, Philippe / Greaves, Ronda / Lackner, Karl J. / Lippi, Giuseppe / Melichar, Bohuslav / Payne, Deborah A.

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Volume 53, Issue 2


Non-invasive prenatal diagnosis of monogenic disorders: an optimized protocol using MEMO qPCR with miniSTR as internal control

Claire Guissart
  • Corresponding author
  • Institut Universitaire de Recherche Clinique, 641 Avenue du Doyen Gaston Giraud, 34093 Montpellier cedex5, France
  • INSERM U827, Université Montpellier I and
  • Laboratoire de Génétique Moléculaire, CHRU Montpellier, Montpellier, France
  • Email
  • Other articles by this author:
  • De Gruyter OnlineGoogle Scholar
/ Vanessa Debant / Marie Desgeorges / Corinne Bareil / Caroline Raynal / Caroline Toga
  • Centre Pluridisciplinaire de Diagnostic Prénatal, Hôpital La Timone, AP-HM, Marseille, France
  • Other articles by this author:
  • De Gruyter OnlineGoogle Scholar
/ Victoria Pritchard / Michel Koenig
  • INSERM U827, Université Montpellier I and
  • Laboratoire de Génétique Moléculaire, CHRU Montpellier, Montpellier, France
  • Other articles by this author:
  • De Gruyter OnlineGoogle Scholar
/ Mireille Claustres
  • INSERM U827, Université Montpellier I and
  • Laboratoire de Génétique Moléculaire, CHRU Montpellier, Montpellier, France
  • Other articles by this author:
  • De Gruyter OnlineGoogle Scholar
/ Marie-Claire Vincent
Published Online: 2014-10-02 | DOI: https://doi.org/10.1515/cclm-2014-0501


Background: Analysis of circulating cell-free fetal DNA (cffDNA) in maternal plasma is very promising for early diagnosis of monogenic diseases. However, this approach is not yet available for routine use and remains technically challenging because of the low concentration of cffDNA, which is swamped by the overwhelming maternal DNA.

Methods: To make clinical applications more readily accessible, we propose a new approach based on mutant enrichment with 3′-modified oligonucleotides (MEMO) PCR along with real-time PCR to selectively amplify from the maternal blood the paternally inherited fetal allele that is not present in the maternal genome.

Results: The first proof of concept of this strategy was displayed for cystic fibrosis by the accuracy of our detection of the p.Gly542* mutation used as the initial developmental model. Subsequently, a retrospective study of plasmas originating from two pregnant women carrying a fetus with private mutation confirmed the effectiveness of our method. We confirmed the presence of cffDNA in the studied samples by the identification of a tri-allelic DNA profile using a miniSTR kit.

Conclusions: This new non-invasive prenatal diagnosis test offers numerous advantages over current methods: it is simple, cost effective, time efficient and does not require complex equipment or bioinformatics settings. Moreover, our assays for different private mutations demonstrate the viability of this approach in clinical settings for monogenic disorders.

Keywords: cell-free fetal DNA; cystic fibrosis; MEMO; non-invasive prenatal diagnosis; quantitative real-time PCR


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About the article

Corresponding author: Dr Claire Guissart, Institut Universitaire de Recherche Clinique, 641 Avenue du Doyen Gaston Giraud, 34093 Montpellier cedex5, France, E-mail: ; INSERM U827, Université Montpellier I and Laboratoire de Génétique Moléculaire, CHRU Montpellier, Montpellier, France

Received: 2014-05-09

Accepted: 2014-09-02

Published Online: 2014-10-02

Published in Print: 2015-02-01

Citation Information: Clinical Chemistry and Laboratory Medicine (CCLM), Volume 53, Issue 2, Pages 205–215, ISSN (Online) 1437-4331, ISSN (Print) 1434-6621, DOI: https://doi.org/10.1515/cclm-2014-0501.

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