Evaluation of the activity of thermostable DNA polymerases in the presence of heme, as a key inhibitor in the real time PCR method in diagnostics of sepsis.

Tomasz Gosiewski; Monika Brzychczy-Włoch; Agata Pietrzyk; Agnieszka Sroka; Małgorzata Bulanda

doi:10.18388/abp.2013_2027

Evaluation of the activity of thermostable DNA polymerases in the presence of heme, as a key inhibitor in the real time PCR method in diagnostics of sepsis.

Tomasz Gosiewski Chair of Microbiology, Jagiellonian University Medical College, Kraków, Poland.;
Monika Brzychczy-Włoch Chair of Microbiology, Jagiellonian University Medical College, Kraków, Poland.;
Agata Pietrzyk Chair of Microbiology, Jagiellonian University Medical College, Kraków, Poland.;
Agnieszka Sroka Chair of Microbiology, Jagiellonian University Medical College, Kraków, Poland.;
Małgorzata Bulanda Chair of Microbiology, Jagiellonian University Medical College, Kraków, Poland.;

DOI: https://doi.org/10.18388/abp.2013_2027

Abstract

The study aim was evaluation of the usefulness of several thermostable DNA polymerases in real time PCR conducted in the presence of the heme. Our study had the advantage of testing several different polymerases, one of which proved to be the least sensitive to heme activity. We also found that there is no need of supplementing the reaction mixture with protective substances like BSA. Selection of the appropriate polymerase can increase the efficiency of the PCR reaction which is very important for diagnosis of sepsis and for other analyses performed on DNA template isolated from the blood.

Published

2013-12-17

Issue

Vol. 60 No. 4 (2013)

Section

Articles

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