MotorPlex provides accurate variant detection across large muscle genes both in single myopathic patients and in pools of DNA samples

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• 作者：Marco Savarese (1) (2)
  Giuseppina Di Fruscio (1) (2)
  Margherita Mutarelli (2)
  Annalaura Torella (1)
  Francesca Magri (3)
  Filippo Maria Santorelli (4)
  Giacomo Pietro Comi (3)
  Claudio Bruno (5)
  Vincenzo Nigro (1) (2)
  
  1. Laboratorio di Genetica Medica ; Dipartimento di Biochimica ; Biofisica e Patologia generale ; Seconda Universit脿 degli Studi di Napoli ; Napoli ; Italy
  2. Telethon Institute of Genetics and Medicine ; Napoli ; Italy
  3. Dino Ferrari Center ; IRCCS Ca鈥?Granda Ospedale Maggiore Policlinico ; Neuroscience Section ; Dipartimento di Fisiopatologia medico-chirurgica e dei trapianti ; Universit脿 di Milano ; Milano ; Italy
  4. Molecular Medicine and Neuromuscular Lab ; IRCCS Stella Maris ; Pisa ; Italy
  5. Centro di Miologia e Patologie Neurodegenerative ; IRCCS Istituto Giannina Gaslini ; Genova ; Italy
  
• 关键词：Next generation sequencing ; Myopathies ; Target sequencing ; Pooling ; Muscular dystrophies
• 刊名：Acta Neuropathologica Communications
• 出版年：2014
• 出版时间：December 2014
• 年：2014
• 卷：2
• 期：1
• 全文大小：451 KB
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• 刊物主题：Neurosciences;
• 出版者：BioMed Central
• ISSN：2051-5960

文摘

Mutations in ~100 genes cause muscle diseases with complex and often unexplained genotype/phenotype correlations. Next-generation sequencing studies identify a greater-than-expected number of genetic variations in the human genome. This suggests that existing clinical monogenic testing systematically miss very relevant information. We have created a core panel of genes that cause all known forms of nonsyndromic muscle disorders (MotorPlex). It comprises 93 loci, among which are the largest and most complex human genes, such as TTN, RYR1, NEB and DMD. MotorPlex captures at least 99.2% of 2,544 exons with a very accurate and uniform coverage. This quality is highlighted by the discovery of 20-30% more variations in comparison with whole exome sequencing. The coverage homogeneity has also made feasible to apply a cost-effective pooled sequencing strategy while maintaining optimal sensitivity and specificity. We studied 177 unresolved cases of myopathies for which the best candidate genes were previously excluded. We have identified known pathogenic variants in 52 patients and potential causative ones in further 56 patients. We have also discovered 23 patients showing multiple true disease-associated variants suggesting complex inheritance. Moreover, we frequently detected other nonsynonymous variants of unknown significance in the largest muscle genes. Cost-effective combinatorial pools of DNA samples were similarly accurate (97-99%). MotorPlex is a very robust platform that overcomes for power, costs, speed, sensitivity and specificity the gene-by-gene strategy. The applicability of pooling makes this tool affordable for the screening of genetic variability of muscle genes also in a larger population. We consider that our strategy can have much broader applications.