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Krzywkowski T, Ciftci S, Assadian F, Nilsson M, Punga T
J. Virol. 91 (11) - [2017-06-01; online 2017-05-12]
An efficient adenovirus infection results in high-level accumulation of viral DNA and mRNAs in the infected cell population. However, the average viral DNA and mRNA content in a heterogeneous cell population does not necessarily reflect the same abundance in individual cells. Here, we describe a novel padlock probe-based rolling-circle amplification technique that enables simultaneous detection and analysis of human adenovirus type 5 (HAdV-5) genomic DNA and virus-encoded mRNAs in individual infected cells. We demonstrate that the method is applicable for detection and quantification of HAdV-5 DNA and mRNAs in short-term infections in human epithelial cells and in long-term infections in human B lymphocytes. Single-cell evaluation of these infections revealed high heterogeneity and unique cell subpopulations defined by differential viral DNA content and mRNA expression. Further, our single-cell analysis shows that the specific expression pattern of viral
PubMed 28298601
DOI 10.1128/JVI.00166-17
Crossref 10.1128/JVI.00166-17
pii: JVI.00166-17
pmc: PMC5432871