Research Papers: Pathology:
Detection and localization of viral infection in the pancreas of patients with type 1 diabetes using short fluorescently-labelled oligonucleotide probes
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Niels Busse1,*, Federico Paroni1,*, Sarah J. Richardson2, Jutta E. Laiho3, Maarit Oikarinen3, Gun Frisk4, Heikki Hyöty3,5, Eelco de Koning6,7, Noel G. Morgan2 and Kathrin Maedler1
1 Islet Biology Laboratory, University of Bremen, Germany
2 Islet Biology Exeter, University of Exeter Medical School, UK
3 Department of Virology, School of Medicine, University of Tampere, Tampere, Finland
4 Department of Immunology, Genetics and Pathology, Uppsala University, Sweden
5 Fimlab Laboratories, Pirkanmaa Hospital District, Tampere, Finland
6 Department of Internal Medicine, Leiden University Medical Center, Leiden, The Netherlands
7 Hubrecht Institute/University Medical Center Utrecht, Utrecht, The Netherlands
* Shared first authors
Federico Paroni, email:
Kathrin Maedler, email:
Keywords: enteroviruses, type 1 diabetes, pancreas, islets, oligonucleotide probes, Pathology Section
Received: November 25, 2016 Accepted: January 19, 2017 Published: January 29, 2017
Enteroviruses, specifically of the Coxsackie B virus family, have been implicated in triggering islet autoimmunity and type 1 diabetes, but their presence in pancreata of patients with diabetes has not been fully confirmed.
To detect the presence of very low copies of the virus genome in tissue samples from T1D patients, we designed a panel of fluorescently labeled oligonucleotide probes, each of 17-22 nucleotides in length with a unique sequence to specifically bind to the enteroviral genome of the picornaviridae family.
With these probes enteroviral RNA was detected with high sensitivity and specificity in infected cells and tissues, including in FFPE pancreas sections from patients with T1D. Detection was not impeded by variations in sample processing and storage thereby overcoming the potential limitations of fragmented RNA. Co-staining of small RNA probes in parallel with classical immunstaining enabled virus detection in a cell-specific manner and more sensitively than by viral protein.
Primary Contact _
Sarah J. Richardson
Jutta E. Laiho
Eelco de Koning
Noel G. Morgan
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