A newborn screening pilot study using methylation-sensitive high resolution melting on dried blood spots to detect Prader-Willi and Angelman syndromes

• Published in: Scientific reports Vol. 10; no. 1; p. 13026
• Main Authors: Ferreira, Igor Ribeiro, Costa, Régis Afonso, Gomes, Leonardo Henrique Ferreira, dos Santos Cunha, Wilton Darleans, Tyszler, Latife Salomão, Freitas, Silvia, Llerena Junior, Juan Clinton, de Vasconcelos, Zilton Farias Meira, Nicholls, Robert D., Guida, Letícia da Cunha
• Format: Journal Article
• Language: English
• Published: London Nature Publishing Group UK 03.08.2020; Nature Publishing Group
• Subjects: 631/1647/2210; 631/208/176; 631/337/176/1968; Angelman Syndrome - blood; Angelman Syndrome - genetics; Autoantigens - genetics; Blood; Deoxyribonucleic acid; DNA; DNA methylation; DNA Methylation - genetics; Dried Blood Spot Testing; Genetic abnormalities; Genetic counseling; Genetic screening; Genomic imprinting; Humanities and Social Sciences; Humans; Infant, Newborn; Medical screening; Melting; Methylation; multidisciplinary; Neonatal Screening; Neonates; Nucleic Acid Denaturation - genetics; Pilot Projects; Prader-Willi syndrome; Prader-Willi Syndrome - blood; Prader-Willi Syndrome - genetics; Ribonuclease P - genetics; Science; Science (multidisciplinary)
• ISSN: 2045-2322; 2045-2322
• DOI: 10.1038/s41598-020-69750-0

Prader-Willi (PWS) and Angelman (AS) syndromes are two clinically distinct imprinted disorders characterized by genetic abnormalities at 15q11-q13. Early diagnosis of both syndromes provides improved treatment and accurate genetic counseling. Whole blood (WB) is the most common DNA source of many methodologies to detect PWS and AS, however, the need of WB makes a massive screening difficult in newborns due to economic and technical limitations. The aim of this study was to adapt a Methylation-sensitive High-Resolution Melting (MS-HRM) approach from dried blood spot (DBS) samples, assessing the different DNA isolation techniques and diagnostic performance. Over a 1-year period, we collected 125 DBS cards, of which 45 had already been diagnosed by MS-HRM (20 PWS, 1 AS, and 24 healthy individuals). We tested three different DBS-DNA extraction techniques assessing the DNA concentration and quality, followed by MS-HRM and statistical comparison. Each DBS-DNA extraction method was capable of accuracy in detecting all PWS and AS individuals. However, the efficiency to detect healthy individuals varied according to methodology. In our experience, DNA extracted from DBS analyzed by the MS-HRM methodology provides an accurate approach for genetic screening of imprinting related disorders in newborns, offering several benefits compared to traditional whole blood methods.