Detection of Mutations in Mitochondrial DNA by Droplet Digital PCR

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J. K. Sofronova¹, Y. Y. Ilinsky^1,2,3, K. E. Orishchenko^1,2, E. G. Chupakhin¹, E. A. Lunev¹, and I. O. Mazunin^1*

¹Immanuil Kant Baltic Federal University, Institute of Chemistry and Biology, 236038 Kaliningrad, Russia; fax: +7 (4012) 595-595; E-mail: IMazunin@kantiana.ru

²Federal Research Center Institute of Cytology and Genetics, Siberian Branch of the Russian Academy of Sciences, 630090 Novosibirsk, Russia; fax: +7 (383) 333-1278

³Novosibirsk State University, 630090 Novosibirsk, Russia; fax: +7 (383) 363-4333

^* To whom correspondence should be addressed.

Received April 22, 2016; Revision received June 13, 2016

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Mutations in mitochondrial DNA (mtDNA) may result in various pathological processes. Detection of mutant mtDNAs is a problem for diagnostic practice that is complicated by heteroplasmy – a phenomenon of the inferring presence of at least two allelic variants of the mitochondrial genome. Also, the level of heteroplasmy largely determines the profile and severity of clinical manifestations. Here we discuss detection of mutations in heteroplasmic mtDNA using up-to-date methods that have not yet been introduced as routine clinical assays. These methods can be used for detecting mutations in mtDNA to verify diagnosis of “mitochondrial disease”, studying dynamics of mutant mtDNA in body tissues of patients, as well as investigating structural features of mtDNAs. Original data on allele-specific discrimination of m.11778G>A mutation by droplet digital PCR are presented, which demonstrate an opportunity for simultaneous detection and quantitative assessment of mutations in mtDNAs.

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KEY WORDS: mitochondria, mutations, mitochondrial diseases, heteroplasmy, droplet digital PCR

DOI: 10.1134/S0006297916100011

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