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Simple, low-cost production of DNA MS2 virus-like particles as molecular diagnostic controls

View ORCID ProfileMichael A. Crone, View ORCID ProfilePaul S. Freemont
doi: https://doi.org/10.1101/2022.09.04.506540
Michael A. Crone
1London Biofoundry, Imperial College Translation and Innovation Hub, White City Campus, 84 Wood Lane, London, W12 0BZ, UK
2Section of Structural and Synthetic Biology, Department of Infectious Disease, Imperial College London, London, SW7 2AZ, UK
3UK Dementia Research Institute Care Research and Technology Centre, Imperial College London, London, UK
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  • ORCID record for Michael A. Crone
Paul S. Freemont
1London Biofoundry, Imperial College Translation and Innovation Hub, White City Campus, 84 Wood Lane, London, W12 0BZ, UK
2Section of Structural and Synthetic Biology, Department of Infectious Disease, Imperial College London, London, SW7 2AZ, UK
3UK Dementia Research Institute Care Research and Technology Centre, Imperial College London, London, UK
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  • For correspondence: p.freemont@imperial.ac.uk
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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 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 MS2 virus-like particles, packaged with DNA, that makes use of affinity chromatography for purification and enzymatic production of exogenous DNA suitable for packaging. The produced virus-like particles 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.

Competing Interest Statement

The authors have declared no competing interest.

Copyright 
The copyright holder for this preprint is the author/funder, who has granted bioRxiv a license to display the preprint in perpetuity. It is made available under a CC-BY 4.0 International license.
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Posted September 04, 2022.
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Simple, low-cost production of DNA MS2 virus-like particles as molecular diagnostic controls
Michael A. Crone, Paul S. Freemont
bioRxiv 2022.09.04.506540; doi: https://doi.org/10.1101/2022.09.04.506540
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Simple, low-cost production of DNA MS2 virus-like particles as molecular diagnostic controls
Michael A. Crone, Paul S. Freemont
bioRxiv 2022.09.04.506540; doi: https://doi.org/10.1101/2022.09.04.506540

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