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Journal Article Ppublish 6 citations

PubMed: 37748509 DOI: 10.1055/a-2153-7789

High Throughput Newborn Screening for Sickle Cell Disease - Application of Two-Tiered Testing with a qPCR-Based Primary screen.

Klin Padiatr 235(6):366-372

Authors: Joachim Janda, Sebastian Hegert, Jessica Bzdok, Rafael Tesorero, Ute Holtkamp, Siegfried Burggraf, Elfriede Schuhmann, Friedrike Hörster, Georg F Hoffmann, Nils Janzen, Jürgen G Okun, Marc Becker, Jürgen Durner

Affiliations

  • Department of General Pediatrics, Division of Neuropediatrics and Metabolic Medicine, University Hospital Heidelberg Center of Paediatric and Adolescent Medicine, Heidelberg, Germany.
  • Screening-Laboratory Hannover, Hannover, Germany.
  • Laboratory Becker & Colleagues, Munich, Germany.
  • Dept. of Clinical Chemistry, Hannover Medical School, Hannover, Germany.
  • Division of Laboratory Medicine, Kinderkrankenhaus auf der Bult, Hannover, Germany.
  • Department of Conservative Dentistry and Periodontology, Ludwig Maximilians University Munich, Munich, Germany.

Abstract

BACKGROUND

Sickle cell disease (SCD) is a group of hemoglobinopathies with a common point mutation causing the production of sickle cell hemoglobin (HbS). In high-throughput newborn screening (NBS) for SCD, a two-step procedure is suitable, in which qPCR first pre-selects relevant samples that are differentiated by a second method.

METHODS

Three NBS centers using qPCR-based primary screening for SCD performed a laboratory comparison. Methods using tandem MS or HPLC were used for differentiation.

RESULTS

In a benchmarking test, 450 dried blood samples were analyzed. Samples containing HbS were detected as reliably by qPCR as by methods established for hemoglobinopathy testing. In a two-step screening approach, the 2-tier-analyses have to distinguish the carrier status from pathological variants. In nine months of regular screening, a total of 353,219 samples were analyzed using two-stage NBS procedures. The 1-tier screening by qPCR reduced the number of samples for subsequent differentiation by>99.5%. Cases with carrier status or other variants were identified as inconspicuous while 78 cases with SCD were revealed. The derived incidence of 1:4,773, is in good agreement with previously published incidences.

CONCLUSION

In high-throughput NBS for SCD, qPCR is suitable to focus 2-tier analyses on samples containing HbS, while being unaffected by factors such as prematurity or transfusions. The substantial reduction of samples numbers positively impacts resource conservation, sustainability, and cost-effectiveness. No false negative cases came to attention.

MeSH

Hemoglobinopathies, Incidence, Anemia, Sickle Cell, Research Design, Pediatrics, Neonatal Screening, Cells, Infant, Newborn, Hemoglobins

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