A Method for Evaluation of the Level of Circulating Mitochondrial DNA by ND1 and ND2 Genes.

Autor: Ogarkov OB; Research Center for Family Health and Human Reproduction Problems, Irkutsk, Russia. obogarkov@mail.ru., Orlova EA; Research Center for Family Health and Human Reproduction Problems, Irkutsk, Russia., Malov IV; Irkutsk State Medical University, Ministry of Health of the Russian Federation, Irkutsk, Russia., Zhdanova SN; Research Center for Family Health and Human Reproduction Problems, Irkutsk, Russia., Malov SI; Irkutsk State Medical University, Ministry of Health of the Russian Federation, Irkutsk, Russia., Khromova PA; Research Center for Family Health and Human Reproduction Problems, Irkutsk, Russia., Stepanenko LA; Irkutsk State Medical University, Ministry of Health of the Russian Federation, Irkutsk, Russia., Sinkov VV; Research Center for Family Health and Human Reproduction Problems, Irkutsk, Russia., Rychkova LV; Research Center for Family Health and Human Reproduction Problems, Irkutsk, Russia., Kolesnikova LI; Research Center for Family Health and Human Reproduction Problems, Irkutsk, Russia.
Jazyk: angličtina
Zdroj: Bulletin of experimental biology and medicine [Bull Exp Biol Med] 2022 Feb; Vol. 172 (4), pp. 495-498. Date of Electronic Publication: 2022 Feb 17.
DOI: 10.1007/s10517-022-05421-6
Abstrakt: The measurement of the level of mitochondrial DNA (mtDNA) in the blood is a difficult problem due to high variability of mitochondrial genes, deletions in the mitochondrial genome in some pathological conditions, different sources of mtDNA into the bloodstream (mtDNA from tissues, from blood cells, etc.). We designed primers and TaqMan probes for highly conserved regions of the ND1 and ND2 genes outside the mitochondrial deletions "hot zones". For standardizing the technique, the true concentration of low-molecular-weight mtDNA was determined by real-time PCR for two targets: a fragment of the ND2 gene (122 bp) and the ND1 and ND2 genes (1198 bp). The sensitivity and specificity of the developed approach were verified on a DNA pool isolated from the blood plasma of healthy donors of various nationalities. The concentration of low-molecular-weight mtDNA in the blood plasma of two patients with COVID-19 was monitored over two weeks of inpatient treatment. A significant increase in the content of low-molecular-weight mtDNA was observed during the first 5 days after hospitalization, followed by a drop to the level of healthy donors. The developed technique makes it possible to assess the blood level of low-molecular-weight mtDNA regardless of the quality of sampling and makes it possible to standardize this biological marker in a wide range of infectious and non-infectious pathologies.
(© 2022. Springer Science+Business Media, LLC, part of Springer Nature.)
Databáze: MEDLINE
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