Simple low-cost production of DNA MS2 virus-like particles as molecular diagnostic controls
File(s)genbio.2022.0033.pdf (800.38 KB)
Published version
Author(s)
Crone, Michael A
Freemont, Paul S
Type
Journal Article
Abstract
Suitable controls are integral for the validation and continued quality assurance of diagnostic workflows. Plasmids, DNA, or in vitro transcribed RNA are often used to validate novel diagnostic workflows, however, they are poorly representative of clinical samples. RNA phage virus-like particles (VLPs) packaged with exogenous RNA have been used in clinical diagnostics as workflow controls, serving as surrogates for infectious viral particles. Comparable controls for DNA viruses are more challenging to produce, with analogous DNA phages being infectious and packaging of DNA within RNA phages requiring complex purification procedures and expensive chemical linkers. We present a simple and inexpensive method to produce Emesvirus zinderi (MS2) VLPs, packaged with DNA, that makes use of affinity chromatography for purification and enzymatic production of exogenous DNA suitable for packaging. The produced VLPs were packaged with hepatitis B virus DNA and were then quantified using droplet digital PCR and calibrated against the WHO international standard using a commercial assay in an accredited clinical laboratory.
Date Issued
2022-12-01
Date Acceptance
2022-10-20
Citation
GEN Biotechnology, 2022, 1 (6), pp.496-503
ISSN
2768-1556
Publisher
Mary Ann Liebert
Start Page
496
End Page
503
Journal / Book Title
GEN Biotechnology
Volume
1
Issue
6
Copyright Statement
© Michael A. Crone and Paul S. Freemont, 2022; Published by Mary Ann Liebert, Inc. This Open Access article is distributed under the terms of the Creative Commons License [CC-BY] ( http://creativecommons.org/licenses/by/4.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.
License URL
Identifier
https://www.ncbi.nlm.nih.gov/pubmed/36644571
PII: 10.1089/genbio.2022.0033
Publication Status
Published
Coverage Spatial
United States