Adjusting MtDNA quantification in whole blood for peripheral blood platelet and leukocyte counts
Entity
UAM. Departamento de Medicina Preventiva y Salud Pública y MicrobiologíaPublisher
Public Library of ScienceDate
2016-10-01Citation
10.1371/journal.pone.0163770
PLOS ONE 11.10 (2016): e016377
ISSN
1932-6203DOI
10.1371/journal.pone.0163770Funded by
This work was supported by Grants PI10/00021 and PI14/00009 (which include FEDER funding) from Instituto de Salud Carlos III supported for research material; FINCyT Science and Technology Program (Scholarships Nº088-FINCyT-BDE-2014 from agreement 1663/OC-PE, between the Republic of Peru and the Inter-American Development Bank) provided support in the form of salaries for author [YHR].Editor's Version
http://dx.doi.org/1932-6203)Subjects
Mitochondrial DNA; Blood; Systemic diseases; Platelet-enriched plasma; Leukocytes; MedicinaRights
© 2016 Hurtado-Roca et al.Abstract
This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.Alterations of mitochondrial DNA copy number (mtDNAcn) in the blood (mitochondrial to nuclear DNA ratio) appear associated with several systemic diseases, including primary mitochondrial disorders, carcinogenesis, and hematologic diseases. Measuring mtDNAcn in DNA extracted from whole blood (WB) instead of from peripheral blood mononuclear cells or buffy coat may yield different results due to mitochondrial DNA present in platelets. The aim of this work is to quantify the contribution of platelets to mtDNAcn in whole blood [mtDNAcn(WB)] and to propose a correction formula to estimate leukocytes' mtDNAcn [mtDNAcn(L)] from mtDNAcn(WB). Blood samples from 10 healthy adults were combined with platelet-enriched plasma and saline solution to produce artificial blood preparations. Aliquots of each sample were combined with five different platelet concentrations. In 46 of these blood preparations, mtDNAcn was measured by qPCR. MtDNAcn(WB) increased 1.07 (95%CI 0.86, 1.29; p<0.001) per 1000 platelets present in the preparation. We proved that leukocyte count should also be taken into account as mtDNAcn(WB) was inversely associated with leukocyte count; it increased 1.10 (95%CI 0.95, 1.25, p<0.001) per unit increase of the ratio between platelet and leukocyte counts. If hematological measurements are available, subtracting 1.10 the platelets/leukocyte ratio from mtDNAcn(WB) may serve as an estimation for mtDNAcn(L). Both platelet and leukocyte counts in the sample are important sources of variation if comparing mtDNAcn among groups of patients when mtDNAcn is measured in DNA extracted from whole blood. Not taking the platelet/leukocyte ratio into account in whole blood measurements, may lead to overestimation and misclassification if interpreted as leukocytes' mtDNAcn.
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Google Scholar:Hurtado-Roca, Yamilee
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Ledesma, Marta
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González-Lázaro, Mónica
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Moreno-Loshuerto, Raquel
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Fernández-Silva, Patricio
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Enriquez, José Antonio
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Laclaustra, Martín
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