Journal
JOURNAL OF VIROLOGICAL METHODS
Volume 178, Issue 1-2, Pages 39-43Publisher
ELSEVIER
DOI: 10.1016/j.jviromet.2011.08.009
Keywords
Q-PCR; Viral safety; Nano-filtration; Parvovirus B19; Virus removal
Funding
- Japanese Human Sciences Foundation [KHB1011]
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The testing of biological products at different stages of the manufacturing process currently involves quantitative polymerase chain reaction (Q-PCR)-based assays. Q-PCR techniques are able to detect not only the viral genome in viral particles but also fragments of degraded genome in samples. The ability of 15 and 19-nm filters to remove viruses was examined by conducting infectivity assays and Q-PCR assays using parvovirus B19 (B19), one of the smallest non-enveloped viruses. Although the filtered samples showed no infectivity, viral DNA was detected by Q-PCR. Interestingly, approximately 90% of the total viral genome in 15-nm filtrates had a detectable size of less than 0.5 kb by the Q-PCR and as a consequence reduction factors were underestimated using Q-PCR. The reduction factors using Q-PCR might be underestimated due to the presence of a large amount of free B19 DNA which shows no infectivity in the tested filtrates. Therefore, the results of Q-PCR should be interpreted with caution. The careful design of primers is needed to eliminate amplification from fragments of viral DNA by Q-PCR. (C) 2011 Elsevier B.V. All rights reserved.
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