Detection and localization of viral infection in the pancreas of patients with type 1 diabetes using short fluorescently-labelled oligonucleotide probes.
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.
Islet Biology Laboratory, University of Bremen, Germany.
Niels
N
Busse
Islet Biology Laboratory, University of Bremen, Germany.
Federico
F
Paroni
Islet Biology Exeter, University of Exeter Medical School, UK.
Sarah J
SJ
Richardson
Department of Virology, School of Medicine, University of Tampere, Tampere, Finland.
Jutta E
JE
Laiho
Department of Virology, School of Medicine, University of Tampere, Tampere, Finland.
Maarit
M
Oikarinen
Department of Immunology, Genetics and Pathology, Uppsala University, Sweden.
Gun
G
Frisk
Department of Virology, School of Medicine, University of Tampere, Tampere, Finland.|Fimlab Laboratories, Pirkanmaa Hospital District, Tampere, Finland.
Heikki
H
Hyöty
Department of Internal Medicine, Leiden University Medical Center, Leiden, The Netherlands.|Hubrecht Institute/University Medical Center Utrecht, Utrecht, The Netherlands.
Eelco
E
de Koning
Islet Biology Exeter, University of Exeter Medical School, UK.
Noel G
NG
Morgan
Islet Biology Laboratory, University of Bremen, Germany.
Kathrin
K
Maedler
D003922:Diabetes Mellitus, Type 1 / Q000821:virology*; D004770:Enterovirus; D004769:Enterovirus Infections / Q000175:diagnosis; D005456:Fluorescent Dyes; D006801:Humans; D007150:Immunohistochemistry; D017404:In Situ Hybridization, Fluorescence / Q000379:methods*; D015345:Oligonucleotide Probes; D010179:Pancreas / Q000821:virology*; D016133:Polymerase Chain Reaction; D012367:RNA, Viral / Q000032:analysis; D012680:Sensitivity and Specificity
D005456:Fluorescent Dyes; D015345:Oligonucleotide Probes; D012367:RNA, Viral
Pathology Section; enteroviruses; islets; oligonucleotide probes; pancreas; type 1 diabetes