Specificity of mtDNA-directed PCR - influence of NUclear MTDNA insertion (NUMT) contamination in routine samples and techniques

• Type: Article in International Scientific Journal
• Date: 2008

Authors

Ana Goios (Author) Other; Lourdes Prieto (Author) Other; Antonio Amorim (Author) FCUP; Luisa Pereira (Author) Other

Journal

Title: International Journal of Legal Medicine

Vol. 122

Pages: 341-345

ISSN: 0937-9827

Publisher: Springer Nature

Indexing

ISI Web of Knowledge - 25 Citations

Scopus - 28 Citations

Scientific classification

FOS: Medical and Health sciences > Other medical sciences

Other information

Authenticus ID: P-003-Y3V

DOI: 10.1007/s00414-007-0191-5

Abstract (EN): Nuclear mitochondrial insertions (NUMTs) are sequences homologous to mtDNA, which are present throughout the human nuclear genome. The possibility that these sequences may be accidentally amplified in reactions directed to mtDNA has been raised and evaluated by different groups and by different means. Despite that, data is still missing on the specificity of PCRs in routine procedures in what concerns contamination with nuclear mtDNA insertions (NUMTs). In this work, we performed PCR sequencing reactions with primers directed either to mitochondrial or to NUMT DNA with different annealing temperatures and in different tissues. We observed that (a) contamination with NUMTs depends on the sample and tissue, and (b) employing routine techniques, there is no risk of co-amplification. Only when mtDNA is almost completely removed from the samples does the number of NUMT copies exceed mitochondrial sequences, i.e., only in samples with virtually no mtDNA, such as those resulting from preferential semen lysis, is there a risk of accidental amplification of NUMTs. We suggest that to evaluate a possible co-amplification of NUMT DNA, it is more relevant to take into account sample processing and original tissue of the samples, and consequently the relative proportions of NUMT and mtDNA, rather than the presence of NUMTs by itself, irrespectively of its proportion.

Language: English

Type (Professor's evaluation): Scientific

No. of pages: 5