Simultaneous detection of multiple viruses in their co-infected cells using multicolour imaging with self-assembled quantum dot probes

The simultaneous detection and evaluation of the coinfection of a cell by multiple viruses or even multiple subtypes still is a difficult challenge. The authors introduce a method for simultaneous imaging, detection and quantitative evaluation of multiple viruses in single cells by using multicolor quantum dot (QD) probes and in a single staining cycle. The multicolor QD probes were fabricated via interaction between QDs conjugated to Staph. aureus protein A (SpA-QDs) and virus-specific antibodies. A cocktail of differently colored QD-SpA-MAbs probes were loaded into the same cells containing multiple viruses, and this enabled the different viruses to be fluorescently imaged and analyzed simultaneously. Specifically, influenza A viruses of type H1N1, H3N2, and H9N2, as well as human adenovirus species B type 3 (HAdV-B3) were imaged and detected in virus-infected cells or in their co-infected cells. In our perception, the method provides a flexible platform for simultaneous detection of multiple viruses in co-infected cells. Hence, it offers new opportunities for the molecular diagnosis of virus coinfection and for studies on virus-cell interactions. [Figure: see text] ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (doi:10.1007/s00604-017-2300-6) contains supplementary material, which is available to authorized users.