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JIMD Reports - Case and Research Reports, 2011/1

Volume 1 of the series JIMD Reports pp 125-129

Date:

Quantitative Analysis of mtDNA Content in Formalin-Fixed Paraffin-Embedded Muscle Tissue

  • Aida FontAffiliated withSección de Errores Congénitos del Metabolismo, Servicio de Bioquímica y Genética Molecular, Hospital Clínic
  • , Frederic TortAffiliated withSección de Errores Congénitos del Metabolismo, Servicio de Bioquímica y Genética Molecular, Hospital ClínicCIBER de Enfermedades Raras (CIBERER)
  • , Aleix Navarro-SastreAffiliated withSección de Errores Congénitos del Metabolismo, Servicio de Bioquímica y Genética Molecular, Hospital ClínicCIBER de Enfermedades Raras (CIBERER)
  • , Victòria CusíAffiliated withServicio de Anatomía Patológica, Hospital San Joan de Déu
  • , Judit García-VilloriaAffiliated withSección de Errores Congénitos del Metabolismo, Servicio de Bioquímica y Genética Molecular, Hospital ClínicCIBER de Enfermedades Raras (CIBERER)
  • , Paz BrionesAffiliated withSección de Errores Congénitos del Metabolismo, Servicio de Bioquímica y Genética Molecular, Hospital ClínicCIBER de Enfermedades Raras (CIBERER)Consejo Superior de Investigaciones Científicas (CSIC)
  • , Antonia RibesAffiliated withSección de Errores Congénitos del Metabolismo, Servicio de Bioquímica y Genética Molecular, Hospital ClínicCIBER de Enfermedades Raras (CIBERER) Email author 

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Abstract

Quantification of mitochondrial DNA (mtDNA) content is an essential tool for the diagnosis of mtDNA depletion syndrome (MDS). Samples collected and processed for anatomopathology studies represent a unique source of archived biological material. Thus, the possibility to study mtDNA copy number in these specimens would be a useful way to screen for MDS. In this study, we designed and validated the methodology to determine mtDNA content by quantitative real-time polymerase chain reaction (qRT-PCR) in formalin-fixed paraffin-embedded (FFPE) muscle tissue. We studied 14 frozen muscle biopsies and compared the results with a portion of the same biopsy embedded in paraffin. Our results showed a similar variability among frozen and FFPE muscle biopsies. Patients with MDS detected in frozen muscle were also confirmed in their corresponding FFPE samples, which validate the usefulness of this approach. We conclude that the analysis of mtDNA copy number in FFPE muscle tissue by qRT-PCR is a useful method for the molecular screening of patients suspected to have MDS when frozen biopsies are not available. Analysis of these samples would facilitate retrospective studies and diagnostic procedures.