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




AuthorsFu, 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

PublisherElsevier BV

Publication year2025

JournalJournal of Clinical Virology

Journal name in sourceJournal of Clinical Virology

Article number105802

Volume178

ISSN1386-6532

eISSN1873-5967

DOIhttps://doi.org/10.1016/j.jcv.2025.105802

Web address https://doi.org/10.1016/j.jcv.2025.105802

Self-archived copy’s web addresshttps://research.utu.fi/converis/portal/detail/Publication/498410249


Abstract

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.


Downloadable publication

This is an electronic reprint of the original article.
This reprint may differ from the original in pagination and typographic detail. Please cite the original version.




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.


Last updated on 2025-11-06 at 11:33