Valiant, W.
, Borman, J.
, Cai, K.
and Vallone, P.
(2023),
Efficient extraction of adventitious virus nucleic acid using commercially available methods, Biologics, [online], https://doi.org/10.1016/j.biologicals.2023.101741, https://tsapps.nist.gov/publication/get_pdf.cfm?pub_id=934925
(Accessed September 17, 2024)
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Efficient extraction of adventitious virus nucleic acid using commercially available methods
Published
Author(s)
, Jon Borman, Kang Cai,
Abstract
An essential step in pharmaceutical product development is screening for contamination with adventitious agents, and there is desire to develop highly sensitive assays to detect adventitious viral nucleic acid. This study sought to examine the nucleic acid extraction efficiency of three viral candidates in relevant background matrices using four different extraction methods. Three model adventitious viruses, Minute virus of Mice, Porcine Circovirus, and Feline Leukemia Virus, were diluted within a variety of background matrices relevant to pharmaceutical production methods. Upon extraction, the nucleic acid was quantified using droplet digital PCR methods. Four nucleic acid extraction methods were assessed, including commercially available kits and manual extraction methods. Each method recovered nucleic acid post-extraction for each of the model viruses within the tested background matrices. The silica-column based method recovered a greater amount of viral nucleic acid, compared to the other methods tested. Similar trends were observed when model virus was diluted in bioreactor supernatant, which replicates industry testing conditions and provides details on which extraction methods might be used in Next Generation Sequencing and PCR methods for detecting contamination within pharmaceutical products.Citation
Biologics
Volume
85
Pub Type
Journals
Download Paper
Keywords
Adventitious viruses, Digital droplet PCR, Nucleic acid extraction, Nuclease, Viral nucleic acid, Contamination
Citation
Issues
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Created December 28, 2023, Updated July 30, 2024