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High-throughput genetic newborn screening for spinal muscular atrophy by rapid nucleic acid extraction from dried blood spots and 384-well qPCR

European Journal of Human Genetics volume 28, pages 23–30 (2020)Cite this article

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Abstract

Establishing nucleic acid-based assays for genetic newborn screening (NBS) provides the possibility to screen for genetically encoded diseases like spinal muscular atrophy (SMA), best before the onset of symptoms. Such assays should be easily scalable to 384-well reactions that make the screening of up to 2000 samples per day possible. We developed a test procedure based on a cleanup protocol for dried blood spots and a quantitative (q)PCR to screen for a homozygous deletion of exon 7 of the survival of motor neuron 1 gene (SMN1) that is responsible for >95% of SMA patients. Performance of this setup is evaluated in detail and tested on routine samples. Our cleanup method for nucleic acids from dried blood spots yields enough DNA for diverse subsequent qPCR applications. To date, we have applied this approach to test 213,279 samples within 18 months. Thirty patients were identified and confirmed, implying an incidence of 1:7109 for the homozygous deletion. Using our cleanup method, a rapid workflow could be established to prepare nucleic acids from dried blood spot cards. Targeting the exon 7 deletion, no invalid, false-positive, or false-negative results were reported to date. This allows timely identification of the disease and grants access to the recently introduced treatment options, in most cases before the onset of symptoms. Carriers are not identified, thus, there are no concerns of whether to report them.

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Acknowledgements

We thank Peta Snikeris for proofing the paper.

Funding

Funding

The screening was funded by the German Cystinosis Foundation (Cystinose Stiftung, DSZ-Regional Office Munich, Widenmayerstr. 10, 80538 Munich, Germany). The funders had no influence on the design, interpretation, and publication of these data.

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Authors and Affiliations

  1. Laboratory Becker & Colleagues, Führichstr. 70, 81671, Munich, Germany

    Ludwig Czibere, Siegfried Burggraf, Tobias Fleige, Birgit Glück, Lisa Marie Keitel, Jürgen Durner, Wulf Röschinger & Marc Becker

  2. TIB Molbiol GmbH, Eresburgstr. 22-23, 12103, Berlin, Germany

    Olfert Landt

  3. Department of Operative/Restorative Dentistry, Periodontology and Pedodontics, Ludwig Maximilian University Munich, Goethestr. 70, 80337, Munich, Germany

    Jürgen Durner & Marc Becker

  4. Pediatric Nephrology, RoMed Klinikum Rosenheim, Pettenkoferstr. 10, 83022, Rosenheim, Germany

    Katharina Hohenfellner

  5. Institute of Human Genetics, University of Cologne, Kerpener Str. 34, 50931, Cologne, Germany

    Brunhilde Wirth

  6. Division of Pediatric Neurology, Center for Neuromuscular Disorders in Childhood. Dr. von Hauner Children’s Hospital, Ludwig-Maximilians-Universität München, Lindwurmstr. 4, 80337, Munich, Germany

    Wolfgang Müller-Felber & Katharina Vill

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  1. Ludwig Czibere
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  2. Siegfried Burggraf
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Correspondence to Ludwig Czibere.

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Czibere, L., Burggraf, S., Fleige, T. et al. High-throughput genetic newborn screening for spinal muscular atrophy by rapid nucleic acid extraction from dried blood spots and 384-well qPCR. Eur J Hum Genet 28, 23–30 (2020). https://doi.org/10.1038/s41431-019-0476-4

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