Effective NPM1 plasmid standards selection for minimal/measurable residual disease monitoring in acute myeloid leukemia.
Acute myeloid leukemia
Cloning
Colony selection
Nucleophosmine 1
Quantitative PCR standards
Journal
Molecular biology reports
ISSN: 1573-4978
Titre abrégé: Mol Biol Rep
Pays: Netherlands
ID NLM: 0403234
Informations de publication
Date de publication:
Aug 2022
Aug 2022
Historique:
received:
03
02
2022
accepted:
10
03
2022
revised:
09
03
2022
pubmed:
19
6
2022
medline:
26
7
2022
entrez:
18
6
2022
Statut:
ppublish
Résumé
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. 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. 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.
Sections du résumé
BACKGROUND
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
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
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.
Identifiants
pubmed: 35716280
doi: 10.1007/s11033-022-07363-8
pii: 10.1007/s11033-022-07363-8
doi:
Substances chimiques
Nuclear Proteins
0
Nucleophosmin
117896-08-9
Types de publication
Journal Article
Langues
eng
Sous-ensembles de citation
IM
Pagination
8169-8172Subventions
Organisme : ministerstvo školství, mládeže a tělovýchovy
ID : MUNI/A/1330/2021
Organisme : ministerstvo zdravotnictví ceské republiky
ID : FNBr, 65269705
Informations de copyright
© 2022. The Author(s), under exclusive licence to Springer Nature B.V.
Références
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