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.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
References
Bai RK, Perng CL, Hsu CH, Wong LJ (2004) Quantitative PCR analysis of mitochondrial DNA content in patients with mitochondrial disease. Ann NY Acad Sci 1011:304–309
Dimmock D, Zhang Q, Dionisi-Vici C et al (2008) Clinical and molecular features of mitochondrial DNA depletion due to mutations in deoxyguanosine kinase. Hum Mutat 29(2):330–331
Dimmock D, Tang L, Schmitt E, Wong L (2010) Quantitative evaluation of the mitochondrial DNA depletion syndrome. Clin Chem 56:1119–1127
Gilbert M, Haselkorn T, Bunce M et al (2007) The isolation of nucleic acids from fixed, paraffin-embedded tissues-which methods are useful and when? PLoS ONE 20(2):e537
Gnanapragasam VJ (2009) Unlocking the molecular archive: the emerging use of formalin-fixed paraffin-embedded tissue for biomarker research in urological cancer. BJU Int 105:274–278
Lehmann U, Kreipe H (2001) Real-Time PCR analysis of DNA and RNA extracted from formalin-fixed and paraffin-embedded biopsies. Methods 25:409–418
Morten KJ, Ashley N, Wijburg F et al (2007) Liver mtDNA content increases during development: a comparison of methods and the importance of age- and tissue-specific controls for the diagnosis of mtDNA depletion. Mitochondrion 7(6):386–395
Rötig A, Poulton J (2009) Genetic causes of mitochondrial DNA depletion in humans. Biochim Biophys Acta 1792:1103–1108
Suomalainen A, Isohanni P (2010) Mitochondrial DNA depletion syndromes-many genes, common mechanisms. Neuromuscul Disord 20:429–437
Yu J, Miller R, Zhang W et al (2008) Copy-number analysis of topoisomerase and thymidylate synthase genes in frozen and FFPE DNAs of colorectal cancers. Pharmacogenomics 9:1459–1466
Acknowledgements
This research was supported in part by the Spanish Ministerio de Sanidad Grant PI08/90348. CIBERER, is an initiative of Instituto de Salud Carlos III.
Author information
Authors and Affiliations
Corresponding author
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2011 SSIEM and Springer-Verlag Berlin Heidelberg
About this chapter
Cite this chapter
Font, A. et al. (2011). Quantitative Analysis of mtDNA Content in Formalin-Fixed Paraffin-Embedded Muscle Tissue. In: JIMD Reports - Case and Research Reports, 2011/1. JIMD Reports, vol 1. Springer, Berlin, Heidelberg. https://doi.org/10.1007/8904_2011_27
Download citation
DOI: https://doi.org/10.1007/8904_2011_27
Received:
Revised:
Accepted:
Published:
Publisher Name: Springer, Berlin, Heidelberg
Print ISBN: 978-3-642-17707-1
Online ISBN: 978-3-642-17708-8
eBook Packages: MedicineMedicine (R0)