Diagnosis of Spinal Muscular Atrophy Via High-Resolution Melting Analysis Symmetric Polymerase Chain Reaction Without Probe: A Screening Evaluation for<i>SMN1</i>Deletions and Intragenic Mutations

Morikawa, Satoru; Harahap, Indra Sari Kusuma; Kaszynski, Richard H.; Yamamoto, Tomoto; Pramudya, Dian Kesuma; Pham, Huyen Thi Van; Hartomo, Tri Budi; Lee, Myeong Jin; Morioka, Ichiro; Nishimura, Noriyuki; Yokoyama, Naoki; Ueno, Yoshio; Matsuo, Masafumi; Nishio, Hisahide

doi:10.1089/gtmb.2010.0237

Cited by 9 publications

(5 citation statements)

References 18 publications

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“…Approximately 95% of SMA patients carry homozygous deletion of the SMN1 gene, and the remaining 5% of the affected individuals may be compound heterozygote for a deleted gene and an intragenic mutation on the other SMN1 gene (Lefebvre et al, 1995;Rodrigues et al, 1995;Kang et al, 2009;Yoon et al, 2010;Morikawa et al, 2011).…”

Section: Introductionmentioning

confidence: 99%

Spinal Muscular Atrophy Carrier Frequency and Estimated Prevalence of the Disease in Moroccan Newborns

Lyahyai

Sbiti

Barkat

et al. 2012

Genetic Testing and Molecular Biomarkers

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Spinal muscular atrophy (SMA) is one of the most common autosomal recessive diseases caused by homozygous deletion of exon 7 of the survival motor neuron 1 (SMN1) gene in approximately 95% of SMA patients. Carrier frequency studies of SMA have been reported for various populations. The aim of our study was to estimate the carrier frequency of the common SMN1 exon 7 deletion in the Moroccan population to achieve an insight into the prevalence of SMA in Morocco. In this study, we used a reliable quantitative real-time polymerase chain reaction assay with SYBR Green I dye to determine the copy number of the SMN1 gene. Analysis of 150 Moroccan newborns predicts a carrier frequency of approximately 1:25, which would mean a calculated SMA prevalence of 1:1800 after correction due to consanguinity. These results show as expected that the SMA carrier frequency in Morocco is higher than in the European populations and is close to those of Middle Eastern countries. Genetic carrier testing for genetic counseling should be recommended particularly to families with a clear clinical history of SMA.

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Section: Introductionmentioning

confidence: 99%

Spinal Muscular Atrophy Carrier Frequency and Estimated Prevalence of the Disease in Moroccan Newborns

Lyahyai

Sbiti

Barkat

et al. 2012

Genetic Testing and Molecular Biomarkers

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“…Two of the melting methods amplify a small amplicon around the informative SNV and can identify many copy number ratios (33,34 ). However, a single copy at each locus (carrier) cannot be distinguished from 2 copies at each locus (normal).…”

Section: Discussionmentioning

confidence: 99%

Copy Number Assessment by Competitive PCR with Limiting Deoxynucleotide Triphosphates and High-Resolution Melting

et al. 2015

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BACKGROUND DNA copy number variation is associated with genetic disorders and cancer. Available methods to discern variation in copy number are typically costly, slow, require specialized equipment, and/or lack precision. METHODS Multiplex PCR with different primer pairs and limiting deoxynucleotide triphosphates (dNTPs) (3–12 μmol/L) were used for relative quantification and copy number assessment. Small PCR products (50–121 bp) were designed with 1 melting domain, well-separated Tms, minimal internal sequence variation, and no common homologs. PCR products were displayed as melting curves on derivative plots and normalized to the reference peak. Different copy numbers of each target clustered together and were grouped by unbiased hierarchical clustering. RESULTS Duplex PCR of a reference gene and a target gene was used to detect copy number variation in chromosomes X, Y, 13, 18, 21, epidermal growth factor receptor (EGFR), survival of motor neuron 1, telomeric (SMN1), and survival of motor neuron 2, centromeric (SMN2). Triplex PCR was used for X and Y and CFTR exons 2 and 3. Blinded studies of 50 potential trisomic samples (13, 18, 21, or normal) and 50 samples with potential sex chromosome abnormalities were concordant to karyotyping, except for 2 samples that were originally mosaics that displayed a single karyotype after growth. Large cystic fibrosis transmembrane conductance regulator (ATP-binding cassette sub-family C, member 7) (CFTR) deletions, EGFR amplifications, and SMN1 and SMN2 copy number assessments were also demonstrated. Under ideal conditions, copy number changes of 1.11-fold or lower could be discerned with CVs of about 1%. CONCLUSIONS Relative quantification by restricting the dNTP concentration with melting curve display is a simple and precise way to assess targeted copy number variation.

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“…4 High-resolution DNA melting analysis (HRMA) HRMA has been used to evaluate individuals for SMA in screening and diagnostic applications. 3,119,120 The difference in SMN1 and SMN2 copy number present in the specimens will create various unique HRMA profiles, distinguishing a heterozygous PCR product from a homozygous product. Therefore, the SMN1 homozygous deletions can be readily identified since these samples will generate the same profile independent of SMN2 copy number.…”

Section: Liquid Microbead Assaysmentioning

confidence: 99%

Newborn screening for spinal muscular atrophy: Anticipating an imminent need

Phan

Taylor

Hannon

et al. 2015

Seminars in Perinatology

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Diagnosis of Spinal Muscular Atrophy Via High-Resolution Melting Analysis Symmetric Polymerase Chain Reaction Without Probe: A Screening Evaluation forSMN1Deletions and Intragenic Mutations

Cited by 9 publications

References 18 publications

Spinal Muscular Atrophy Carrier Frequency and Estimated Prevalence of the Disease in Moroccan Newborns

Spinal Muscular Atrophy Carrier Frequency and Estimated Prevalence of the Disease in Moroccan Newborns

Copy Number Assessment by Competitive PCR with Limiting Deoxynucleotide Triphosphates and High-Resolution Melting

Newborn screening for spinal muscular atrophy: Anticipating an imminent need

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