DNA sequencing of maternal plasma to identify Down syndrome and other trisomies in multiple gestations

Authors

  • Jacob A. Canick,

    Corresponding author
    • Division of Medical Screening and Special Testing, Department of Pathology and Laboratory Medicine, Women and Infants Hospital, Alpert Medical School of Brown University, Providence, RI, USA
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  • Edward M. Kloza,

    1. Division of Medical Screening and Special Testing, Department of Pathology and Laboratory Medicine, Women and Infants Hospital, Alpert Medical School of Brown University, Providence, RI, USA
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  • Geralyn M. Lambert-Messerlian,

    1. Division of Medical Screening and Special Testing, Department of Pathology and Laboratory Medicine, Women and Infants Hospital, Alpert Medical School of Brown University, Providence, RI, USA
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  • James E. Haddow,

    1. Division of Medical Screening and Special Testing, Department of Pathology and Laboratory Medicine, Women and Infants Hospital, Alpert Medical School of Brown University, Providence, RI, USA
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  • Mathias Ehrich,

    1. Sequenom, Inc., San Diego, CA, USA
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  • Dirk van den Boom,

    1. Sequenom, Inc., San Diego, CA, USA
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  • Allan T. Bombard,

    1. Sequenom, Inc., San Diego, CA, USA
    2. Sequenom Center for Molecular Medicine, San Diego, CA, USA
    3. Department of Reproductive Medicine, University of California San Diego, San Diego, CA, USA
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  • Cosmin Deciu,

    1. Sequenom Center for Molecular Medicine, San Diego, CA, USA
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  • Glenn E. Palomaki

    1. Division of Medical Screening and Special Testing, Department of Pathology and Laboratory Medicine, Women and Infants Hospital, Alpert Medical School of Brown University, Providence, RI, USA
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  • This article was published online on May 14, 2012. Errors were subsequently identified in the Abstract. This notice is included in the online and print versions to indicate that both have been corrected [June 6, 2012].

  • Funding sources: Sequenom, Inc. fully funded the project through a grant to Women & Infants Hospital of Rhode Island. Sequenom Center for Molecular Medicine (SCMM) was responsible for developing an internally validated laboratory developed test (LDT) for detecting Down syndrome in maternal plasma using massively parallel shotgun sequencing and for providing clinical interpretation of the test results. SCMM also identified, equipped and trained an independent laboratory to test a subset of samples through a separate contract with UCLA. The sponsor did not control study design, identify or communicate with Enrollment Sites, thaw or test samples prior to the formal testing period, have access to patient information prior to all testing being completed, analyze study results, prepare drafts of the manuscript, or have final decisions on manuscript content.

  • Conflicts of interest: Palomaki and Canick were members of the Sequenom Clinical Advisory Board for 6 months, and resigned when the study was funded in 2008. Van den Boom, Ehrich, and Bombard are employees and shareholders of Sequenom, Inc. Deciu is an employee of Sequenom Center for Molecular Medicine and a shareholder of Sequenom, Inc.

Jacob A. Canick. E-mail: jcanick@wihri.org

ABSTRACT

Objective

Studies on prenatal testing for Down syndrome (trisomy 21), trisomy 18, and trisomy 13 by massively parallel shotgun sequencing (MPSS) of circulating cell free DNA have been, for the most part, limited to singleton pregnancies. If MPSS testing is offered clinically, it is important to know if these trisomies will also be identified in multiple pregnancies.

Method

Among a cohort of 4664 high-risk pregnancies, maternal plasma samples were tested from 25 twin pregnancies (17 euploid, five discordant and two concordant for Down syndrome; one discordant for trisomy 13) and two euploid triplet pregnancies [Correction made here after initial online publication.]. Results were corrected for GC content bias. For each target chromosome (21, 18, and 13), z-scores of 3 or higher were considered consistent with trisomy.

Results

Seven twin pregnancies with Down syndrome, one with trisomy 13, and all 17 twin euploid pregnancies were correctly classified [detection rate 100%, 95% confidence interval (CI) 59%–100%, false positive rate 0%, 95% CI 0%–19.5%], as were the two triplet euploid pregnancies.

Conclusion

Although study size is limited, the underlying biology combined with the present data provide evidence that MPSS testing can be reliably used as a secondary screening test for Down syndrome in women with high-risk twin gestations. © 2012 John Wiley & Sons, Ltd.

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