Spinal muscular atrophy genotyping by gene dosage using multiple ligation-dependent probe amplification

Scarciolla, Oronzo; Stuppia, Liborio; Angelis, Maria Vittoria De; Murru, S; Palka, Chiara; Giuliani, Rossella; Pace, Marta; Muzio, Antonio Di; Torrente, Isabella; Morella, Annunziata; Grammatico, Paola; Giacanelli, Manlio; Rosatelli, C; Uncini, Antonino; Dallapiccola, Bruno

doi:10.1007/s10048-006-0051-3

Cited by 64 publications

(46 citation statements)

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“…Reports indicate that the MLPA assay is better and more efficient than the 2-step DHPLC. The MLPA assay provides the unique ability to hybridize several probes specific for SMN1 and SMN2 genes in a single experiment [9].…”

mentioning

confidence: 99%

Determination of SMN1 and SMN2 Copy Numbers in a Korean Population using Multiplex Ligation-dependent Probe Amplification

Yoon

Lee

2010

Ann Lab Med

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mentioning

confidence: 99%

Determination of SMN1 and SMN2 Copy Numbers in a Korean Population using Multiplex Ligation-dependent Probe Amplification

Yoon

Lee

2010

Ann Lab Med

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“…Исходя из этого, можно предположить, что увеличение копий гена SMN2, приводит к повышению экспрессии полноценного белка SMN с центромерных копий гена на геном и тем мягче течение заболевания [13].…”

Section: Discussionunclassified

Mutational analysis of SMN gene in patients with spinal muscular atrophy and the correlation between mutation and the severity of clinical manifestations

et al. 2017

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Objective: studying of mutations in the SMN1, SMN2 and NAIP genes in the diagnosis of spinal muscular atrophy and determining the correlation between the mutation and the severity of clinical manifestations.Methods: 82 children with suspicion of spinal muscular atrophy were examined. Genomic DNA was isolated from the whole blood of patients using the Wizard®Genomic DNA Purification kit (Promega). Further diagnostics was carried out using the MLPA method on a 96-capillary automatic analyzer 3730xl DNA Analyzer. The results were processed using the software Coffalyser DB.v.130.Results: A homozygous deletion of the exons 7-8 of the SMN1 gene was observed in all SMA patients of this study. At the same time, the deletion of the exon 7 of the SMN1 gene and an increase in the number of copies of SMN2, 1 (2%) and 2 (4%), respectively, were found among patients with type II and III type of SMA. In the analysis of deletions of exons 7-8 of the SMN1 gene, the SMN1 gene was completely deleted in 15 (29%), 2 (4%) and 1 (2%) SMA I, II and III patients.The study revealed that the NAIP gene was deleted in 8 patients (19.2%) with type I CMA and in two patients (4%) with SMA type II. Also, in a single patient with a type II SMA, deletions of the exon 8 of the SMN1 1 gene (2%) was found.Conclusion: the MLPA method allows not only to confirm the diagnosis of spinal amyotrophy, but also to specify the type of spinal amyotrophy depending on the detected changes in the SMN1, SMN2 and NAIP genes. The clinical diagnosis of SMA is mainly based on the identification of homozygous deletions of the 7th and 8th exons of the SMN1 gene. Deletion of the NAIP gene and an increase in the number of copies of SMN2 are considered to assess the clinical phenotype of CMA patients.Keywords: spinal muscular atrophy, genes SMN1, SMN2 and NAIP, MLPA -method, deletion. Зерттеу мақсаты: Спинальды бұлшықетті атрофиясының диагностикасында (СБА) SMN1, SMN2 және NAIP гендеріндегі мутациялар-ды зерттеу және мутация мен клиникалық көріністердің ауырлығы арасындағыбайланыстыанықтау.Әдістері:Спинальдыбұлшықетті атрофия күдікпен 82 бала тексерілді. Genomic DNA Wizard® Genomic DNA Purification Kit (Promega) жиынтық көмегімен пациенттердің тұтас қанынан геномдық ДНҚ бөлектелген. Әрі қарай, 96 капиллярлық автоматты 3730xl DNA Analyser анализатор арқылы MLPA әдісімен диагностика өткізілді. Қорытындылар Coffalyser DB.v.130 бағдарламалық жасақтама арқылы өңделді.Нәтижелері: Өткізілген зерттеуде барлық пациенттерде SMN1 геннің7-8 экзондарында гомозиготтық делеция анықталды. Бес паци-ентте СБА 1 типімен SMN1 геннің7 экзонда гомозиготтық делеция анықталды. Сонымен қатар, СБА II және III типтері бар науқастардың арасында SMN1 генінің 7 экзонының делециясы және SMN2 ген генінің көшірмелерінің көбеюі анықталды, 1 (2%) және 2 (4%), тиісінше. SMN1 генінің 7-8 экзондарын делециясын талдау кезінде, I, II және III типтермен СБА науқастарда 15 (29%), 2 (4%) және 1 (2%) SMN1 генде толықтай делеция көрсетілген.

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“…The number of SMN2 copies (arranged in tandem in cis configuration on each chromosome) ranges from zero to five that can be detected using quantitative PCR and MLPA methods [8,9]. The presence of three or more copies of SMN2 is associated with a milder phenotype [10][11][12][13].…”

Section: Copy Number Variation Of Smn2 Genementioning

confidence: 99%

Mutation Spectrum of Survival Motor Neuron Gene in Spinal Muscular Atrophy

Rad¹

2017

Down Syndr Chr Abnorm

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Clinical Aspects of SMASpinal muscular atrophy (SMA), with an incidence of 1/10000 world-wide [1,2] is a lower motor neuron disease and characterized by symmetrical muscle weakness and atrophy resulting from progressive degeneration of alpha motor neurons and loss of the anterior horn cells in the spinal cord and the brain stem nuclei. The inheritance pattern of SMA is autosomal recessive and classified clinically into the following types, based on the age at onset and clinical severity: SMA 0 (prenatal onset), SMA I (Werdnig-Hoffmann disease with onset before age 6 months), SMA II (Dubowitz disease with onset between age 6 and 18 months), SMA III (Kugelberg-Welander disease with onset after age 18 months), and SMA IV (onset after age 18 years). After unraveling the genetic basis of SMA, all SMN1-associated clinical types can be considered as a continuum without a clear delineation. Survival Motor Neuron (SMN) GeneSurvival motor neuron (SMN) gene, the SMA determining gene, located on 5q12.2-q13.3 with two homologue copies, SMN1 (survival motor neuron 1; telomeric copy) and SMN2 (survival motor neuron 2; centromeric copy), which differ by only eight nucleotide (five are intronic and three are exonic, located within exons 6, 7, and 8) [3][4][5][6]. SMN1 and SMN2, both containing nine exons with 99% nucleotide identity, are arranged in tandem on each chromosome encoding a 294-amino acid RNA-binding protein. One of the coding sequence of SMN2 that differs from that of SMN1 by a single nucleotide (840 C>T) results in alternative splicing of exon 7 [4]. Due to the alternative splicing of exon 7, SMN2 genes produce a reduced amount of full length transcripts, and a variable amount of mRNA lacking exon 7, which give raise to a truncated and unstable protein [7], whereas SMN1 produces full-length transcripts.Full-length product of SMN1 is necessary for lower motor neuron function and loss of SMN1 is essential to the pathogenesis of SMA, while SMN2 copy number modifies the severity of phenotype. Copy Number Variation of SMN2 GeneThe number of SMN2 copies (arranged in tandem in cis configuration on each chromosome) ranges from zero to five that can be detected using quantitative PCR and MLPA methods [8,9]. The presence of three or more copies of SMN2 is associated with a milder phenotype [10][11][12][13]. All SMA patients retain at least one copy of SMN2, so, SMA is caused by low levels of SMN protein rather than complete absence of the protein. Most SMA type I patients have two copies of SMN2, three SMN2 copies are common in SMA type II, while type III and IV generally have three or four copies. On the other hand, no correlation exists between the loss of SMN1 exon 7 and the severity of disease, that is, the homozygous exon 7 deletions is observed with the same frequency in all phenotypes. Mutation Spectrum of SMN1 GeneApproximately 95%-98% of all types of SMA patients show homozygous deletion of SMN1 exon 7 (and exon 8 in the majority of cases) [14][15][16]. Approximately 6% of parents of a child with homozygous SMN1 del...

show abstract

Spinal muscular atrophy genotyping by gene dosage using multiple ligation-dependent probe amplification

Cited by 64 publications

References 22 publications

Determination of SMN1 and SMN2 Copy Numbers in a Korean Population using Multiplex Ligation-dependent Probe Amplification

Determination of SMN1 and SMN2 Copy Numbers in a Korean Population using Multiplex Ligation-dependent Probe Amplification

Mutational analysis of SMN gene in patients with spinal muscular atrophy and the correlation between mutation and the severity of clinical manifestations

Mutation Spectrum of Survival Motor Neuron Gene in Spinal Muscular Atrophy

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