A1 Refereed original research article in a scientific journal
Use of polyethylene glycol precipitation and ultracentrifugation to enhance the sensitivity of hepatitis B virus DNA detection
Authors: Fu, Michael X.; Larralde, Osmany; Mayne, Richard; Kean, Kai; Reid, Kaitlin; Andersson, Monique; Golubchik, Tanya; McKeating, Jane A.; Jarvis, Lisa; Irving, William L.; Simmonds, Peter; Harvala, Heli
Publisher: Elsevier BV
Publication year: 2025
Journal: Journal of Clinical Virology
Journal name in source: Journal of Clinical Virology
Article number: 105802
Volume: 178
ISSN: 1386-6532
eISSN: 1873-5967
DOI: https://doi.org/10.1016/j.jcv.2025.105802
Web address : https://doi.org/10.1016/j.jcv.2025.105802
Self-archived copy’s web address: https://research.utu.fi/converis/portal/detail/Publication/498410249
Background: Sensitive molecular detection of hepatitis B virus (HBV) DNA is crucial for diagnosing and managing occult hepatitis. To improve the sensitivity of HBV DNA detection, we compared the effectiveness of polyethylene glycol (PEG) precipitation and ultracentrifugation to concentrate DNA prior to extraction.
Methods: Twenty-three HBV DNA-positive samples with low viral loads were compared between the extraction of standard (0.2 mL) and larger volumes (5 mL) of plasma, through PEG precipitation of 10 mL and 20 mL of plasma, and ultracentrifugation from 35 mL of plasma. The effectiveness of the methods for HBV DNA detection was assayed by quantitative PCR. For genetic characterisation, Sanger sequencing of amplicons and targeted Illumina sequencing were used. Costs, sample capacities, and turnaround times were compared.
Results: DNA was detected in a greater number of samples using PEG and ultracentrifugation (detecting up to all 23 samples) compared to more standard extraction methods (detecting at least 18 samples). Efficiencies of recovery of HBV DNA from samples were comparable in all concentration methods. HBV and other DNA viruses, such as human herpesviruses and anelloviruses, were detected in samples and at higher read counts with PEG concentration than without. The availability, cost, relative simplicity, and throughput of PEG precipitation conferred further advantages to ultracentrifugation.
Conclusions: PEG precipitation from large volumes of plasma is a practical and economical alternative to ultracentrifugation and could be a similarly effective concentration method for low viral load samples in blood donation and clinical virology laboratories.
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Funding information in the publication:
This work and authors [Michael X Fu, Richard Mayne, Kai Kean, Kaitlin Reid, Tanya Golubchik, Peter Simmonds, Heli Harvala] was supported by the National Institutes for Health and Care Research [grant number NIHR203338]. The funding body had no role in the study's design, data collection, analysis, or manuscript writing.