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

被引:25
作者
Cote, Helene C. F. [1 ]
Gerschenson, Mariana [2 ,3 ]
Walker, Ulrich A. [4 ,5 ]
Miro, Oscar [6 ]
Garrabou, Gloria [6 ]
Hammond, Emma [7 ]
Villarroya, Joan [8 ,9 ]
Giralt, Marta [8 ,9 ]
Villarroya, Francesc [8 ,9 ]
Cinque, Paola [10 ]
Garcia-Arumi, Elena [11 ]
Andreu, Antonio L. [11 ]
Pinti, Marcello [12 ]
Cossarizza, Andrea [12 ]
机构
[1] Univ British Columbia, Dept Pathol & Lab Med, Vancouver, BC V6T 2B5, Canada
[2] Univ Hawaii Manoa, Dept Cell & Mol Biol, Honolulu, HI 96822 USA
[3] Univ Hawaii Manoa, Hawaii Ctr AIDS, Honolulu, HI 96822 USA
[4] Univ Basel, Dept Rheumatol, CH-4003 Basel, Switzerland
[5] Univ Freiburg, Dept Rheumatol, D-7800 Freiburg, Germany
[6] Univ Barcelona, Hosp Clin Barcelona, Dept Internal Med, Mitochondrial Res Lab,IDIBAPS, Barcelona, Spain
[7] Murdoch Univ, Perth, WA, Australia
[8] Inst Salud Carlos III, CIBER Fisiopatol Obesidad & Nutr, Barcelona, Spain
[9] Univ Barcelona, Dept Biochem & Mol Biol, Barcelona, Spain
[10] Ist Sci San Raffaele, I-20132 Milan, Italy
[11] Univ Autonoma Barcelona, Univ Vall dHebron, Inst Recerca VHIR, Unitat Patol Mitocondrial,CIBBIM,CIBERER ISCIII, E-08193 Barcelona, Spain
[12] Univ Modena & Reggio Emilia, Dept Biomed Sci, I-41125 Modena, Italy
基金
美国国家卫生研究院;
关键词
Inter-laboratory variability; mtDNA content by qPCR; NUCLEOSIDE ANALOGS; GENE-EXPRESSION; HUMAN GENOME; CELL-LINES; THERAPY; LYMPHOCYTES; PSEUDOGENES; DEPLETION; DISEASE; CANCER;
D O I
10.1016/j.mito.2011.01.011
中图分类号
Q2 [细胞生物学];
学科分类号
071009 ; 090102 ;
摘要
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 R-2=0.664; logio-transformed data R-2=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. (C) 2011 Elsevier B.V. and Mitochondria Research Society. All rights reserved.
引用
收藏
页码:520 / 527
页数:8
相关论文
共 27 条
  • [1] Quantification of mitochondrial DNA copy number: Pre-analytical factors
    Andreu, Antonio L.
    Martinez, Ramiro
    Marti, Ramon
    Garcia-Arumi, Elena
    [J]. MITOCHONDRION, 2009, 9 (04) : 242 - 246
  • [2] Mitochondrial pseudogenes: evolution's misplaced witnesses
    Bensasson, D
    Zhang, DX
    Hartl, DL
    Hewitt, GM
    [J]. TRENDS IN ECOLOGY & EVOLUTION, 2001, 16 (06) : 314 - 321
  • [3] Gene duplication: A drive for phenotypic diversity and cause of human disease
    Conrad, Bernard
    Antonarakis, Stylianos E.
    [J]. ANNUAL REVIEW OF GENOMICS AND HUMAN GENETICS, 2007, 8 : 17 - 35
  • [4] Mutational and selective effects on copy-number variants in the human genome
    Cooper, Gregory M.
    Nickerson, Deborah A.
    Eichler, Evan E.
    [J]. NATURE GENETICS, 2007, 39 (Suppl 7) : S22 - S29
  • [5] Inherited mitochondrial diseases of DNA replication
    Copeland, William C.
    [J]. ANNUAL REVIEW OF MEDICINE, 2008, 59 : 131 - 146
  • [6] Cossarizza A, 2003, ANTIVIR THER, V8, P315
  • [7] Côte HCF, 2005, ANTIVIR THER, V10, pM3
  • [8] Perinatal exposure to antiretroviral therapy is associated with increased blood mitochondrial DNA levels and decreased mitochondrial gene expression in infants
    Cote, Helene C. F.
    Raboud, Janet
    Bitnun, Ari
    Alimenti, Ariane
    Money, Deborah M.
    Maan, Evelyn
    Costei, Adriana
    Gadawski, Izabelle
    Diong, Christina
    Read, Stanley
    Shen, Sandy
    Harrigan, P. Richard
    Burdge, David R.
    King, Susan M.
    Forbes, John C.
    [J]. JOURNAL OF INFECTIOUS DISEASES, 2008, 198 (06): : 851 - 859
  • [9] Cottier M, 2004, INT J MOL MED, V14, P483
  • [10] Galluzzi L, 2005, ANTIVIR THER, V10, P191