Wijaya, Yogik Onky Silvana and Nishio, Hisahide and Niba, Emma Tabe Eko and Shiroshita, Tomoyoshi and Kato, Masako and Bouike, Yoshihiro and Tode, Chisato and Ar Rochmah, Mawaddah and Harahap, Nur Imma Fatimah and Nurputra, Dian Kesumapramudya and Okamoto, Kentaro and Saito, Toshio and Takeuchi, Atsuko and Lai, Poh San and Yamaguchi, Seiji and Shinohara, Masakazu (2021) Dried Blood Spot Screening System for Spinal Muscular Atrophy with Allele-Specific Polymerase Chain Reaction and Melting Peak Analysis. Genetic Testing and Molecular Biomarkers, 25 (4). 293 – 301. ISSN 19450265
Full text not available from this repository. (Request a copy)Abstract
Background and Aim: Spinal muscular atrophy (SMA) is a lower motor neuron disease with autosomal recessive inheritance caused by homozygous SMN1 deletions. Although SMA has been considered as incurable, newly developed drugs improve life prognoses and motor functions of patients. To maximize the efficacy of the drugs, SMA patients should be treated before symptoms become apparent. Thus, newborn screening for SMA is strongly recommended. In this study, we aim to establish a new simple screening system based on DNA melting peak analysis. Materials and Methods: A total of 124 dried blood spot (DBS) on FTA® ELUTE cards (51 SMN1-deleted patients with SMA, 20 carriers, and 53 controls) were punched and subjected to direct amplification of SMN1 and CFTR (reference gene). Melting peak analyses were performed to detect SMN1 deletions from DBS samples. Results: A combination of allele-specific polymerase chain reaction (PCR) and melting peak analyses clearly distinguished the DBS samples with and without SMN1. Compared with the results of fresh blood samples, our new system yielded 100 sensitivity and specificity. The advantages of our system include (1) biosafe collection, transfer, and storage for DBS samples, (2) obviating the need for DNA extraction from DBS preventing contamination, (3) preclusion of fluorescent probes leading to low PCR cost, and (4) fast and high-throughput screening for SMN1 deletions. Conclusion: We demonstrate that our system would be applicable to a real-world newborn screening program for SMA, because our new technology is efficient for use in routine clinical laboratories that do not have highly advanced PCR instruments. © Copyright 2021, Mary Ann Liebert, Inc., publishers 2021.
Item Type: | Article |
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Additional Information: | Cited by: 5 |
Uncontrolled Keywords: | Cystic Fibrosis Transmembrane Conductance Regulator; DNA; Dried Blood Spot Testing; Exons; Female; Gene Deletion; Gene Frequency; High-Throughput Screening Assays; Humans; Infant, Newborn; Male; Muscular Atrophy, Spinal; Neonatal Screening; Nucleic Acid Denaturation; Real-Time Polymerase Chain Reaction; Sensitivity and Specificity; Survival of Motor Neuron 1 Protein; cystic fibrosis transmembrane conductance regulator; DNA; fluorescent dye; survival motor neuron protein 1; CFTR protein, human; cystic fibrosis transmembrane conductance regulator; DNA; SMN1 protein, human; survival motor neuron protein 1; allele specific polymerase chain reaction; Article; blood sampling; contamination; controlled study; DNA denaturation; DNA extraction; dried blood spot testing; gene amplification; gene deletion; high throughput screening; human; major clinical study; sensitivity and specificity; spinal muscular atrophy; storage; blood; DNA denaturation; dried blood spot testing; exon; female; gene frequency; genetics; male; metabolism; newborn; newborn screening; procedures; real time polymerase chain reaction; spinal muscular atrophy |
Subjects: | R Medicine > RB Biomedical Sciences |
Divisions: | Faculty of Medicine, Public Health and Nursing > Public Health and Nutrition |
Depositing User: | Sri JUNANDI |
Date Deposited: | 25 Sep 2024 06:52 |
Last Modified: | 25 Sep 2024 06:52 |
URI: | https://ir.lib.ugm.ac.id/id/eprint/4651 |