Universität zu Köln

Czibere, Ludwig ORCID: 0000-0002-7508-5841, Burggraf, Siegfried ORCID: 0000-0002-2116-0156, Fleige, Tobias, Glueck, Birgit, Keitel, Lisa Marie, Landt, Olfert, Durner, Juergen, Roeschinger, Wulf, Hohenfellner, Katharina, Wirth, Brunhilde ORCID: 0000-0003-4051-5191, Mueller-Felber, Wolfgang, Vill, Katharina and Becker, Marc (2020). 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 (1). S. 23 - 31. LONDON: NATURE PUBLISHING GROUP. ISSN 1476-5438

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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.

Item Type:	Journal Article
Creators:	Creators Email ORCID ORCID Put Code Czibere, Ludwig UNSPECIFIED orcid.org/0000-0002-7508-5841 UNSPECIFIED Burggraf, Siegfried UNSPECIFIED orcid.org/0000-0002-2116-0156 UNSPECIFIED Fleige, Tobias UNSPECIFIED UNSPECIFIED UNSPECIFIED Glueck, Birgit UNSPECIFIED UNSPECIFIED UNSPECIFIED Keitel, Lisa Marie UNSPECIFIED UNSPECIFIED UNSPECIFIED Landt, Olfert UNSPECIFIED UNSPECIFIED UNSPECIFIED Durner, Juergen UNSPECIFIED UNSPECIFIED UNSPECIFIED Roeschinger, Wulf UNSPECIFIED UNSPECIFIED UNSPECIFIED Hohenfellner, Katharina UNSPECIFIED UNSPECIFIED UNSPECIFIED Wirth, Brunhilde UNSPECIFIED orcid.org/0000-0003-4051-5191 UNSPECIFIED Mueller-Felber, Wolfgang UNSPECIFIED UNSPECIFIED UNSPECIFIED Vill, Katharina UNSPECIFIED UNSPECIFIED UNSPECIFIED Becker, Marc UNSPECIFIED UNSPECIFIED UNSPECIFIED
URN:	urn:nbn:de:hbz:38-352634
DOI:	10.1038/s41431-019-0476-4
Journal or Publication Title:	Eur. J. Hum. Genet.
Volume:	28
Number:	1
Page Range:	S. 23 - 31
Date:	2020
Publisher:	NATURE PUBLISHING GROUP
Place of Publication:	LONDON
ISSN:	1476-5438
Language:	English
Faculty:	Unspecified
Divisions:	Unspecified
Subjects:	no entry
Uncontrolled Keywords:	Keywords Language DIAGNOSIS; SMN1; IDENTIFICATION; NUSINERSEN; CHALLENGES; MANAGEMENT; PCR Multiple languages Biochemistry & Molecular Biology; Genetics & Heredity Multiple languages
URI:	http://kups.ub.uni-koeln.de/id/eprint/35263