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.
| Original language | English |
|---|---|
| Pages (from-to) | 520-527 |
| Number of pages | 8 |
| Journal | Mitochondrion |
| Volume | 11 |
| Issue number | 3 |
| DOIs | |
| Publication status | Published - May 2011 |
Fingerprint
Keywords
- Inter-laboratory variability
- MtDNA content by qPCR
ASJC Scopus subject areas
- Cell Biology
- Molecular Biology
- Molecular Medicine
Cite this
Quality assessment of human mitochondrial DNA quantification : MITONAUTS, an international multicentre survey. / Côté, Hélène C F; Gerschenson, Mariana; Walker, Ulrich A.; Miro, Oscar; Garrabou, Gloria; Hammond, Emma; Villarroya, Joan; Giralt, Marta; Villarroya, Francesc; Cinque, Paola; Garcia-Arumi, Elena; Andreu, Antonio L.; Pinti, Marcello; Cossarizza, Andrea.
In: Mitochondrion, Vol. 11, No. 3, 05.2011, p. 520-527.Research output: Contribution to journal › Article
}
TY - JOUR
T1 - Quality assessment of human mitochondrial DNA quantification
T2 - MITONAUTS, an international multicentre survey
AU - Côté, Hélène C F
AU - Gerschenson, Mariana
AU - Walker, Ulrich A.
AU - Miro, Oscar
AU - Garrabou, Gloria
AU - Hammond, Emma
AU - Villarroya, Joan
AU - Giralt, Marta
AU - Villarroya, Francesc
AU - Cinque, Paola
AU - Garcia-Arumi, Elena
AU - Andreu, Antonio L.
AU - Pinti, Marcello
AU - Cossarizza, Andrea
PY - 2011/5
Y1 - 2011/5
N2 - 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.
AB - 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.
KW - Inter-laboratory variability
KW - MtDNA content by qPCR
UR - http://www.scopus.com/inward/record.url?scp=79953755706&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=79953755706&partnerID=8YFLogxK
U2 - 10.1016/j.mito.2011.01.011
DO - 10.1016/j.mito.2011.01.011
M3 - Article
C2 - 21303702
AN - SCOPUS:79953755706
VL - 11
SP - 520
EP - 527
JO - Mitochondrion
JF - Mitochondrion
SN - 1567-7249
IS - 3
ER -