Quality assessment of human mitochondrial DNA quantification: MITONAUTS, an international multicentre survey

Hélène C.F. Côté, Mariana Gerschenson, Ulrich A. Walker, Oscar Miro, Gloria Garrabou, Emma Hammond, Joan Villarroya, Marta Giralt, Francesc Villarroya, Paola Cinque, Elena Garcia-Arumi, Antonio L. Andreu, Marcello Pinti, Andrea Cossarizza

    Research output: Contribution to journalArticleResearchpeer-review

    19 Citations (Scopus)

    Abstract

    Mitochondrial DNA quantification by qPCR is used in the context of many diseases and toxicity studies but comparison of results between laboratories is challenging. Through two multigroup distributions of DNA samples from human cell lines, the MITONAUTS group anonymously compared mtDNA/nDNA quantification across nine laboratories involved in HIV research worldwide. Eight of the nine sites showed significant correlation between them (mean raw data R2=0.664; log10-transformed data R2=0.844). Although mtDNA/nDNA values were well correlated between sites, the inter-site variability on the absolute measurements remained high with a mean (range) coefficient of variation of 71 (37-212) %. Some variability appeared cell line-specific, probably due to chromosomal alterations or pseudogenes affecting the quantification of certain genes, while within cell line variability was likely due to differences in calibration of the standard curves. The use of two mtDNA and two single copy nDNA genes with highly specific primers to quantify each genome would help address copy number variants. Our results indicate that sample shipment must be done frozen and that absolute mtDNA/nDNA ratio values cannot readily be compared between laboratories, especially if assessing cultured cell mtDNA content. However, within laboratory and relative mtDNA/nDNA comparisons between laboratories should be reliable. © 2011 © Elsevier B.V. and Mitochondria Research Society. All rights reserved.
    Original languageEnglish
    Pages (from-to)520-527
    JournalMitochondrion
    Volume11
    Issue number3
    DOIs
    Publication statusPublished - 1 May 2011

    Keywords

    • Inter-laboratory variability
    • MtDNA content by qPCR

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