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Clinical Chemistry and Laboratory Medicine (CCLM)

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

Editor-in-Chief: Plebani, Mario / Plebani, Mario

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


IMPACT FACTOR 2018: 3.638

CiteScore 2018: 2.44

SCImago Journal Rank (SJR) 2018: 1.191
Source Normalized Impact per Paper (SNIP) 2018: 1.205

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1437-4331
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Volume 53, Issue 2

Issues

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

Abstract

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

References

  • 1.

    Tsui NB, Lo YM. Recent advances in the analysis of fetal nucleic acids in maternal plasma. Curr Opin Hematol 2012;19:462–8.Web of SciencePubMedCrossrefGoogle Scholar

  • 2.

    Rodríguez de Alba M, Bustamante-Aragonés A, Perlado S, Trujillo-Tiebas MJ, Díaz-Recasens J, Plaza-Arranz J, et al. Noninvasive prenatal diagnosis of monogenic disorders. Expert Opin Biol Ther 2012;12(Suppl 1):S171–9.CrossrefWeb of ScienceGoogle Scholar

  • 3.

    Bustamante-Aragonés A, Rodríguez de Alba M, Perlado S, Trujillo-Tiebas MJ, Arranz JP, Díaz-Recasens J, et al. Non-invasive prenatal diagnosis of single-gene disorders from maternal blood. Gene 2012;504:144–9.CrossrefWeb of ScienceGoogle Scholar

  • 4.

    Chan KC. Clinical applications of the latest molecular diagnostics in noninvasive prenatal diagnosis. Top Curr Chem 2014;336: 47–65.PubMedCrossrefGoogle Scholar

  • 5.

    Bustamante-Aragones A, Gonzalez-Gonzalez C, de Alba MR, Ainse E, Ramos C. Noninvasive prenatal diagnosis using ccffDNA in maternal blood: state of the art. Expert Rev Mol Diagn 2010;10:197–205.Web of ScienceCrossrefGoogle Scholar

  • 6.

    Mujezinovic F, Alfirevic Z. Procedure-related complications of amniocentesis and chorionic villous sampling: a systematic review. Obstet Gynecol 2007;110:687–94.CrossrefWeb of SciencePubMedGoogle Scholar

  • 7.

    Lo YM, Corbetta N, Chamberlain PF, Rai V, Sargent IL, Redman CW, et al. Presence of fetal DNA in maternal plasma and serum. Lancet 1997;350:485–7.PubMedCrossrefGoogle Scholar

  • 8.

    Honda H, Miharu N, Ohashi Y, Samura O, Kinutani M, Hara T, et al. Fetal gender determination in early pregnancy through qualitative and quantitative analysis of fetal DNA in maternal serum. Hum Genet 2002;110:75–9.PubMedCrossrefGoogle Scholar

  • 9.

    Alberry M, Maddocks D, Jones M, Abdel Hadi M, Abdel-Fattah S, Avent N, et al. Free fetal DNA in maternal plasma in anembryonic pregnancies: confirmation that the origin is the trophoblast. Prenat Diagn 2007;27:415–8.PubMedCrossrefWeb of ScienceGoogle Scholar

  • 10.

    Chan KC, Zhang J, Hui AB, Wong N, Lau TK, Leung TN, et al. Size distributions of maternal and fetal DNA in maternal plasma. Clin Chem 2004;50:88–92.CrossrefPubMedGoogle Scholar

  • 11.

    Lun FM, Chiu RW, Allen Chan KC, Yeung Leung T, Kin Lau T, Dennis Lo YM. Microfluidics digital PCR reveals a higher than expected fraction of fetal DNA in maternal plasma. Clin Chem 2008;54:1664–72.CrossrefPubMedWeb of ScienceGoogle Scholar

  • 12.

    Lo YM, Tein MS, Lau TK, Haines CJ, Leung TN, Poon PM, et al. Quantitative analysis of fetal DNA in maternal plasma and serum: implications for noninvasive prenatal diagnosis. Am J Hum Genet 1998;62:768–75.PubMedCrossrefGoogle Scholar

  • 13.

    Lo YM, Zhang J, Leung TN, Lau TK, Chang AM, Hjelm NM. Rapid clearance of fetal DNA from maternal plasma. Am J Hum Genet 1999;64:218–24.PubMedCrossrefGoogle Scholar

  • 14.

    Lench N, Barrett A, Fielding S, McKay F, Hill M, Jenkins L, et al. The clinical implementation of non-invasive prenatal diagnosis for single-gene disorders: challenges and progress made. Prenat Diagn 2013;33:555–62.PubMedWeb of ScienceCrossrefGoogle Scholar

  • 15.

    Lee S-T, Kim J-Y, Kown M-J, Kim SW, Chung JH, Ahn M-J, et al. Mutant enrichment with 3’-modified oligonucleotides a practical PCR method for detecting trace mutant DNAs. J Mol Diagn 2011;13:657–68.Web of ScienceCrossrefGoogle Scholar

  • 16.

    Salvatore D, Buzzetti R, Baldo E, Forneris MP, Lucidi V, Manunza D, et al. An overview of international literature from cystic fibrosis registries. Part 3. Disease incidence, genotype/phenotype correlation, microbiology, pregnancy, clinical complications, lung transplantation, and miscellanea. J Cyst Fibros 2011;10: 71–85.Web of SciencePubMedCrossrefGoogle Scholar

  • 17.

    Claustres M, Guittard C, Bozon D, Chevalier F, Verlingue C, Ferec C, et al. Spectrum of CFTR mutations in cystic fibrosis and in congenital absence of the vas deferens in France. Hum Mutat 2000;16:143–56.PubMedCrossrefGoogle Scholar

  • 18.

    Bustamante-Aragones A, Gallego-Merlo J, Trujillo-Tiebas MJ, de Alba MR, Gonzalez-Gonzalez C, Glover G, et al. New strategy for the prenatal detection/exclusion of paternal cystic fibrosis mutations in maternal plasma. J Cyst Fibros 2008;7:505–10.Web of ScienceCrossrefGoogle Scholar

  • 19.

    González-González MC, García-Hoyos M, Trujillo MJ, Rodríguez de Alba M, Lorda-Sánchez I, Díaz-Recasens J, et al. Prenatal detection of a cystic fibrosis mutation in fetal DNA from maternal plasma. Prenat Diagn 2002;22:946–8.CrossrefGoogle Scholar

  • 20.

    Nasis O, Thompson S, Hong T, Sherwood M, Radcliffe S, Jackson L, et al. Improvement in sensitivity of allele-specific PCR facilitates reliable noninvasive prenatal detection of cystic fibrosis. Clin Chem 2004;50:694–701.CrossrefPubMedGoogle Scholar

  • 21.

    Wright CF, Burton H. The use of cell-free fetal nucleic acids in maternal blood for non-invasive prenatal diagnosis. Hum Reprod Update 2009;15:139–51.Web of SciencePubMedCrossrefGoogle Scholar

  • 22.

    Grubwieser P, Mühlmann R, Berger B, Niederstätter H, Pavlic M, Parson W. A new “miniSTR-multiplex” displaying reduced amplicon lengths for the analysis of degraded DNA. Int J Legal Med 2006;120:115–20.PubMedCrossrefGoogle Scholar

  • 23.

    Milbury CA, Li J, Makrigiorgos GM. COLD-PCR-enhanced high-resolution melting enables rapid and selective identification of low-level unknown mutations. Clin Chem 2009;55:2130–43.PubMedCrossrefWeb of ScienceGoogle Scholar

  • 24.

    Galbiati S, Brisci A, Damin F, Gentilin B, Curcio C, Restagno G, et al. Fetal DNA in maternal plasma: a noninvasive tool for prenatal diagnosis of beta-thalassemia. Expert Opin Biol Ther 2012;12(Suppl 1):S181–7.Web of ScienceCrossrefGoogle Scholar

  • 25.

    Macher HC, Martinez-Broca MA, Rubio-Calvo A, Leon-Garcia C, Conde-Sanchez M, Costa A, et al. Non-invasive prenatal diagnosis of multiple endocrine neoplasia type 2A using COLD-PCR combined with HRM genotyping analysis from maternal serum. PLoS One 2012;7:e51024.Web of ScienceCrossrefGoogle Scholar

  • 26.

    Phylipsen M, Yamsri S, Treffers EE, Jansen DT, Kanhai WA, Boon EM, et al. Non-invasive prenatal diagnosis of beta-thalassemia and sickle-cell disease using pyrophosphorolysis-activated polymerization and melting curve analysis. Prenat Diagn 2012;32:578–87.PubMedWeb of ScienceCrossrefGoogle Scholar

  • 27.

    Page-Christiaens GC, Bossers B, van der Schoot CE, De Haas M. Use of bi-allelic insertion/deletion polymorphisms as a positive control for fetal genotyping in maternal blood: first clinical experience. Ann N Y Acad Sci 2006;1075:123–9.Google Scholar

  • 28.

    Doescher A, Petershofen EK, Wagner FF, Schunter M, Müller TH. Evaluation of single-nucleotide polymorphisms as internal controls in prenatal diagnosis of fetal blood groups. Transfusion 2013;53:353–62.Web of SciencePubMedCrossrefGoogle Scholar

  • 29.

    Mulero JJ, Chang CW, Lagacé RE, Wang DY, Bas JL, McMahon TP, et al. Development and validation of the AmpFlSTR MiniFiler PCR Amplification Kit: a MiniSTR multiplex for the analysis of degraded and/or PCR inhibited DNA. J Forensic Sci 2008;53:838–52.CrossrefPubMedGoogle Scholar

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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