Effective NPM1 plasmid standards selection for minimal/measurable residual disease monitoring in acute myeloid leukemia.

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Authors

FOLTA Adam SASINKOVA Markéta ĎURINÍKOVÁ Anna DRNCOVÁ Marie WEINBERGEROVÁ Barbora MAYER Jiří JEŽÍŠKOVÁ Ivana

Year of publication 2022
Type Article in Periodical
Magazine / Source Molecular Biology Reports
MU Faculty or unit

Faculty of Medicine

Citation
Web https://link.springer.com/article/10.1007/s11033-022-07363-8
Doi http://dx.doi.org/10.1007/s11033-022-07363-8
Keywords Nucleophosmine 1; Acute myeloid leukemia; Quantitative PCR standards; Colony selection; Cloning
Description Background NPM1 plasmid standards are required for absolute quantification of minimal residual disease in acute myeloid leukemia patients. The standards are usually obtained, next to commercially constructed gene fragments, from transgenic bacteria colonies. However, this procedure is laborious and very time consuming. Methods and results We have developed a PCR method that speeds up, simplifies, and streamlines the process of preparing NPM1 plasmid standards. The method is based on a combination of three primers, two surrounding the usual NPM1 mutation position and one over the mutation site. With this method, we were able to clearly distinguish plasmids with at least 15 different NPM1 mutations from the wild-type NPM1 plasmid. Conclusions With the new approach, preparing NPM1 plasmid standards is easier, identifying NPM1-positive colonies is possible in less than a day and moreover, for a lower price than commercially constructed gene fragments.
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