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 : EJHG
Ludwig CzibereMarc Becker

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 a...Continue Reading

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Citations

Oct 2, 2019·European Journal of Human Genetics : EJHG·Tobias FleigeMarc Becker
Dec 5, 2019·Developmental Medicine and Child Neurology·Karolina Aragon-GawinskaUNKNOWN SMA-Registry Study Group
Oct 9, 2019·Journal of Neuromuscular Diseases·Katharina VillWolfgang Müller-Felber
May 19, 2020·Genetics in Medicine : Official Journal of the American College of Medical Genetics·Denise M KayMichele Caggana
May 18, 2020·Journal of Neuromuscular Diseases·Kyriaki KekouJoanne Traeger-Synodinos
Apr 29, 2020·Diagnostics·Kristina MalsagovaAnna Kaysheva
Mar 8, 2020·Journal of Neuromuscular Diseases·Wolfgang Müller-FelberHeike Kölbel
Jul 17, 2020·Journal of Clinical Medicine·Sonia Messina, Maria Sframeli
Oct 20, 2020·International Journal of Neonatal Screening·Masakazu ShinoharaUNKNOWN SMA-NBS PILOT STUDY GROUP
Oct 20, 2020·International Journal of Neonatal Screening·Yogik Onky Silvana WijayaMasakazu Shinohara
Dec 29, 2020·Degenerative Neurological and Neuromuscular Disease·Maria Jędrzejowska
Feb 10, 2021·International Journal of Neonatal Screening·Yogik Onky Silvana WijayaMasakazu Shinohara
Apr 4, 2021·International Journal of Neonatal Screening·Katerina S KuceraLisa M Gehtland
Apr 21, 2021·Genetic Testing and Molecular Biomarkers·Yogik Onky Silvana WijayaMasakazu Shinohara
Jun 15, 2021·Frontiers in Genetics·Ziga I RemecMarusa Debeljak

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Methods Mentioned

BETA
SMA
PCR
nucleic acid extraction
antisense oligonucleotides

Software Mentioned

LightCycler
MedCalc

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