High-resolution melting (HRM) analysis as a feasible method for detecting spinal muscular atrophy via dried blood spots

2012 ◽  
Vol 413 (21-22) ◽  
pp. 1781-1785 ◽  
Author(s):  
Tze-Kiong Er ◽  
Tzu-Min Kan ◽  
Yu-Fa Su ◽  
Ta-Chih Liu ◽  
Jan-Gowth Chang ◽  
...  
Keyword(s):  
High Resolution ◽  
Spinal Muscular Atrophy ◽  
High Resolution Melting ◽  
Muscular Atrophy ◽  
Dried Blood Spots ◽  
Feasible Method ◽  
Blood Spots ◽  
Hrm Analysis

scholarly journals Newborn Screening for Spinal Muscular Atrophy by Calibrated Short-Amplicon Melt Profiling

2012 ◽  
Vol 58 (6) ◽  
pp. 1033-1039 ◽  
Author(s):  
Steven F Dobrowolski ◽  
Ha T Pham ◽  
Frances Pouch Downes ◽  
Thomas W Prior ◽  
Edwin W Naylor ◽  
...  
Keyword(s):  
Spinal Muscular Atrophy ◽  
Newborn Screening ◽  
Motor Neuron ◽  
Muscular Atrophy ◽  
Dried Blood Spots ◽  
Continuous Treatment ◽  
Management Options ◽  
Blood Spots ◽  
Survival Of Motor Neuron ◽  
Homozygous Deletions

Abstract BACKGROUND The management options for the autosomal recessive neurodegenerative disorder spinal muscular atrophy (SMA) are evolving; however, their efficacy may require presymptom diagnosis and continuous treatment. To identify presymptomatic SMA patients, we created a DNA-based newborn screening assay to identify the homozygous deletions of the SMN1 (survival of motor neuron 1, telomeric) gene observed in 95%–98% of affected patients. METHODS We developed primers that amplify a 52-bp PCR product from homologous regions in the SMN1 and SMN2 (survival of motor neuron 2, centromeric) genes that flank a divergent site at site c.840. Post-PCR high-resolution melt profiling assessed the amplification product, and we used a unique means of melt calibration to normalize profiles. Samples that we had previously characterized for the numbers of SMN1 and SMN2 copies established genotypes associated with particular profiles. The system was evaluated with approximately 1000 purified DNA samples, 100 self-created dried blood spots, and >1200 dried blood spots from newborn screening tests. RESULTS Homozygous deletion of SMN1 exon 7 produced a distinctive melt profile that identified SMA patients. Samples with different numbers of SMN1 and SMN2 copies were resolved by their profiles. All samples with homozygous deletions were unambiguously recognized, and no normal sample was misidentified as a positive. CONCLUSIONS This assay has characteristics suitable for population-based screening. A reliable screening test will facilitate the identification of an SMA-affected cohort to receive early intervention to maximize the benefit from treatment. A prospective screening trial will allow the efficacy of treatment options to be assessed, which may justify the inclusion of SMA as a target for population screening.

scholarly journals Rapid diagnosis of spinal muscular atrophy using High-Resolution Melting Analysis

2009 ◽  
Vol 10 (1) ◽  
Author(s):  
Wan Jin Chen ◽  
Wan Juan Dong ◽  
Xiao Zhen Lin ◽  
Min Ting Lin ◽  
Shen Xing Murong ◽  
...  
Keyword(s):  
High Resolution ◽  
Spinal Muscular Atrophy ◽  
High Resolution Melting ◽  
Muscular Atrophy ◽  
Rapid Diagnosis ◽  
High Resolution Melting Analysis ◽  
Melting Analysis

scholarly journals Assessment of Spinal Muscular Atrophy Carrier Status by Determining SMN1 Copy Number Using Dried Blood Spots

2020 ◽  
Vol 6 (2) ◽  
pp. 43
Author(s):  
Yogik Onky Silvana Wijaya ◽  
Jamiyan Purevsuren ◽  
Nur Imma Fatimah Harahap ◽  
Emma Tabe Eko Niba ◽  
Yoshihiro Bouike ◽  
...  
Keyword(s):  
Spinal Muscular Atrophy ◽  
Copy Number ◽  
Muscular Atrophy ◽  
Autosomal Recessive Inheritance ◽  
Dried Blood Spots ◽  
Carrier Screening ◽  
Carrier Status ◽  
Essential Information ◽  
Blood Spots ◽  
Incurable Disease

Spinal muscular atrophy (SMA) is a common neuromuscular disease with autosomal recessive inheritance. The disease gene, SMN1, is homozygously deleted in 95% of SMA patients. Although SMA has been an incurable disease, treatment in infancy with newly developed drugs has dramatically improved the disease severity. Thus, there is a strong rationale for newborn and carrier screening for SMA, although implementing SMA carrier screening in the general population is controversial. We previously developed a simple, accurate newborn SMA screening system to detect homozygous SMN1 deletions using dried blood spots (DBS) on filter paper. Here, we modified our previous system to detect the heterozygous deletions of SMN1, which indicates SMA carrier status. The system involves a calibrator-normalized relative quantification method using quantitative nested PCR technology. Our system clearly separated the DBS samples with one SMN1 copy (carrier status with a heterozygous deletion of SMN1) from the DBS samples with two SMN1 copies (non-carrier status with no deletion of SMN1). We also analyzed DBS samples from SMA families, confirmed SMA in the affected children, and determined the carrier status of their parents based on the SMN1 copy number. In conclusion, our system will provide essential information for risk assessment and genetic counseling, at least for SMA families.

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

2020 ◽  
Vol 10 (1) ◽  
Author(s):  
Igor Ribeiro Ferreira ◽  
Régis Afonso Costa ◽  
Leonardo Henrique Ferreira Gomes ◽  
Wilton Darleans dos Santos Cunha ◽  
Latife Salomão Tyszler ◽  
...  
Keyword(s):  
Pilot Study ◽  
High Resolution ◽  
Newborn Screening ◽  
High Resolution Melting ◽  
Dried Blood Spots ◽  
Blood Spots
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