Detection and differentiation of murine leukemia virus (MLV) and murine stem cell virus (MSCV) and therefrom derived nucleic acids.
Gene transfer
MoMLV
MuLV
Provirus
Virus titration
Journal
Journal of virological methods
ISSN: 1879-0984
Titre abrégé: J Virol Methods
Pays: Netherlands
ID NLM: 8005839
Informations de publication
Date de publication:
01 2022
01 2022
Historique:
received:
07
07
2021
revised:
01
10
2021
accepted:
03
10
2021
pubmed:
11
10
2021
medline:
22
3
2022
entrez:
10
10
2021
Statut:
ppublish
Résumé
Murine leukemia virus (MLV) and murine stem cell virus (MSCV) and derived retroviral vectors are widely used to study retrovirus biology and as tools for gene delivery. The method described here represents a quantitative real time PCR (qPCR) with hydrolysis probe that can be applied within classical qPCR as well as in digital droplet PCR (ddPCR). The method targets a 60 bp long fragment located within the U5 region of the MLV/MSCV genome sequence. For the here described method a LOD95% of 25 copies per PCR reaction (DNA) and 80 copies per PCR reaction (RNA) was determined, and PCR efficiencies of 92.5 % and 98.5 %, respectively, were observed. This method enables the fast and simple titration of viral genomic RNA present in retroviral vector stocks for accurate and consistent transduction experiments. Furthermore, it enables the detection of proviral and transfer plasmid derived DNA sequences and can be modified to differentiate between retroviral RNA and DNA.
Identifiants
pubmed: 34627947
pii: S0166-0934(21)00255-X
doi: 10.1016/j.jviromet.2021.114316
pii:
doi:
Substances chimiques
Nucleic Acids
0
Types de publication
Journal Article
Research Support, Non-U.S. Gov't
Langues
eng
Sous-ensembles de citation
IM
Pagination
114316Informations de copyright
Copyright © 2021 Elsevier B.V. All rights reserved.