Application of real-time PCR to significantly reduce the time to obtain recombinant MVA virus

•Novel single-tube real-time PCR to determine a ratio of wild type and rMVA in plaques.•Rapid generation of new pure rMVA in 11 days.•Plaque purification is less effective then end-point dilution as the last purification step.•Various marker gene selected plaques differ greatly in proportion of rMVA...

Full description

Saved in:

Bibliographic Details
Published in:	Journal of virological methods Vol. 289; p. 114056
Main Authors:	Orlova, O.V., Glazkova, D.V., Tsyganova, G.M., Antoshkina, I.V., Mintaev, R.R., Tikhonov, A.S., Bogoslovskaya, E.V., Shipulin, G.A.
Format:	Journal Article
Language:	English
Published:	Netherlands Elsevier B.V 01-03-2021
Subjects:	Animals Antigens, Viral Cell Line Genetic Vectors Humans MVA Real-time PCR Real-Time Polymerase Chain Reaction SARS-CoV-2 SARS-CoV-2 - genetics Vaccine Vaccinia virus - isolation & purification MVA SARS-CoV-2 Vaccine Real-time PCR
Online Access:	Get full text
Tags:	Add Tag No Tags, Be the first to tag this record!

Description
Summary:	•Novel single-tube real-time PCR to determine a ratio of wild type and rMVA in plaques.•Rapid generation of new pure rMVA in 11 days.•Plaque purification is less effective then end-point dilution as the last purification step.•Various marker gene selected plaques differ greatly in proportion of rMVA. Obtaining a pure recombinant Modified Vaccinia Ankara (MVA) virus is a multistage, time-consuming procedure. We describe a novel single-tube real-time PCR which enables determination of the amount of wild type and recombinant viruses and their ratio in plaques. Use of the real-time PCR significantly reduce the time and efforts needed to obtain purified recombinant MVA. The new approach has been applied to generate recombinant MVAs encoding different SARS-COV-2 antigens.
Bibliography:	ObjectType-Article-1 SourceType-Scholarly Journals-1 ObjectType-Feature-2 content type line 23
ISSN:	0166-0934 1879-0984
DOI:	10.1016/j.jviromet.2020.114056