Detection and localization of viral infection in the pancreas of patients with type 1 diabetes using short fluorescently-labelled oligonucleotide probes

Oncotarget
Niels BusseKathrin Maedler

Abstract

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.

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Citations

Dec 4, 2019·Nature Medicine·Kendra VehikUNKNOWN TEDDY Study Group
Sep 20, 2019·Nature Reviews. Endocrinology·Jorma IlonenRiitta Veijola
Jul 12, 2020·Microorganisms·Shirin GeravandiKathrin Maedler
Jan 11, 2021·Clinica Chimica Acta; International Journal of Clinical Chemistry·Feliciana Real-FernándezAnna Maria Papini
Dec 29, 2020·Clinical and Experimental Immunology·M BattagliaT I Tree
Jun 29, 2021·Frontiers in Immunology·James Alexander PearsonLi Wen

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Methods Mentioned

BETA
PCR
Assay
fluorescence microscopy
biopsy
biopsies
FCS
PCRs

Software Mentioned

ClustalW
ImageJ
Exeter
Quantigene ® ViewRNA
Stellaris RNA FISH Probe Designer

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