Journal
VACCINE
Volume 37, Issue 7, Pages 1001-1005Publisher
ELSEVIER SCI LTD
DOI: 10.1016/j.vaccine.2018.12.019
Keywords
Adventitious agent; Virus; Degenerate PCR; High throughput sequencing (HTS)
Categories
Funding
- FDA/CBER intramural research program
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We used a combination of DOP-PCR with high throughput sequencing (HTS) to study infected cell cultures over time to assess the feasibility of using this technique to provide a read-out other than cytopathic effect in cell culture infectivity assays. Because DOP-PCR primers feature a short constant sequence at their 3' terminus, the procedure yields a reproducible representational library of products from a given PCR template, including viral nucleic acids. Using SV40- and MVM-infected cultures harvested at different times, we show that the number of viral matches among DOP-PCR products parallels the quantity of virus as shown by real-time PCR, and further show that HTS analysis of specific DOP-PCR products that increase in quantity over time could be used to identify the infecting virus with a sensitivity similar to that of typical cell-culture assays that rely on cytopathic effect. Published by Elsevier Ltd.
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