Comprehensive Mutation Analysis for Congenital Muscular Dystrophy: A Clinical PCR-Based Enrichment and Next-Generation Sequencing Panel

Research output: Contribution to journalArticle


  • C. Alexander Valencia
  • Arunkanth Ankala
  • Devin Rhodenizer
  • Shruti Bhide
  • Martin Robert Littlejohn
  • Lisa Mari Keong
  • Anne Rutkowski
  • Susan Sparks
  • Carsten Bonnemann
  • Madhuri Hegde

External Institution(s)

  • Emory University
  • Cincinnati Children's Hospital Medical Center
  • University of Cincinnati
  • University of California at Los Angeles
  • Carolinas Medical Center
  • National Institutes of Health


Original languageEnglish (US)
Article numbere53083
JournalPloS one
Issue number1
StatusPublished - Jan 17 2013


The congenital muscular dystrophies (CMDs) comprise a heterogeneous group of heritable muscle disorders with often difficult to interpret muscle pathology, making them challenging to diagnose. Serial Sanger sequencing of suspected CMD genes, while the current molecular diagnostic method of choice, can be slow and expensive. A comprehensive panel test for simultaneous screening of mutations in all known CMD-associated genes would be a more effective diagnostic strategy. Thus, the CMDs are a model disorder group for development and validation of next-generation sequencing (NGS) strategies for diagnostic and clinical care applications. Using a highly multiplexed PCR-based target enrichment method (RainDance) in conjunction with NGS, we performed mutation detection in all CMD genes of 26 samples and compared the results with Sanger sequencing. The RainDance NGS panel showed great consistency in coverage depth, on-target efficiency, versatility of mutation detection, and genotype concordance with Sanger sequencing, demonstrating the test's appropriateness for clinical use. Compared to single tests, a higher diagnostic yield was observed by panel implementation. The panel's limitation is the amplification failure of select gene-specific exons which require Sanger sequencing for test completion. Successful validation and application of the CMD NGS panel to improve the diagnostic yield in a clinical laboratory was shown.

Citation formats



Valencia, CA, Ankala, A, Rhodenizer, D, Bhide, S, Littlejohn, MR, Keong, LM, Rutkowski, A, Sparks, S, Bonnemann, C & Hegde, M 2013, 'Comprehensive Mutation Analysis for Congenital Muscular Dystrophy: A Clinical PCR-Based Enrichment and Next-Generation Sequencing Panel', PloS one, vol. 8, no. 1, e53083.