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

Niels Busse; Federico Paroni; Sarah J Richardson; Jutta E Laiho; Maarit Oikarinen; Gun Frisk; Heikki Hyöty; Eelco de Koning; Noel G Morgan; Kathrin Maedler

doi:10.18632/oncotarget.14896

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

Niels Busse, Federico Paroni, Sarah J Richardson, Jutta E Laiho, Maarit Oikarinen, Gun Frisk, Heikki Hyöty, Eelco de Koning, Noel G Morgan, Kathrin Maedler

Hubrecht Institute

Research output: Contribution to journal/periodical › Article › Scientific › peer-review

25 Citations (Scopus)

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.

Original language	English
Pages (from-to)	12620-12636
Number of pages	17
Journal	Oncotarget
Volume	8
Issue number	8
DOIs	https://doi.org/10.18632/oncotarget.14896
Publication status	Published - 21 Feb 2017

Keywords

Diabetes Mellitus, Type 1
Enterovirus
Enterovirus Infections
Fluorescent Dyes
Humans
Immunohistochemistry
In Situ Hybridization, Fluorescence
Oligonucleotide Probes
Pancreas
Polymerase Chain Reaction
RNA, Viral
Sensitivity and Specificity
Journal Article

Access to Document

10.18632/oncotarget.14896

Cite this

Busse, N., Paroni, F., Richardson, S. J., Laiho, J. E., Oikarinen, M., Frisk, G., Hyöty, H., de Koning, E., Morgan, N. G., & Maedler, K. (2017). Detection and localization of viral infection in the pancreas of patients with type 1 diabetes using short fluorescently-labelled oligonucleotide probes. Oncotarget, 8(8), 12620-12636. https://doi.org/10.18632/oncotarget.14896

@article{28251823ddc749bba34e05af4445543c,

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

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.",

keywords = "Diabetes Mellitus, Type 1, Enterovirus, Enterovirus Infections, Fluorescent Dyes, Humans, Immunohistochemistry, In Situ Hybridization, Fluorescence, Oligonucleotide Probes, Pancreas, Polymerase Chain Reaction, RNA, Viral, Sensitivity and Specificity, Journal Article",

author = "Niels Busse and Federico Paroni and Richardson, {Sarah J} and Laiho, {Jutta E} and Maarit Oikarinen and Gun Frisk and Heikki Hy{\"o}ty and {de Koning}, Eelco and Morgan, {Noel G} and Kathrin Maedler",

year = "2017",

month = feb,

day = "21",

doi = "10.18632/oncotarget.14896",

language = "English",

volume = "8",

pages = "12620--12636",

journal = "Oncotarget",

issn = "1949-2553",

publisher = "Impact Journals",

number = "8",

}

TY - JOUR

T1 - Detection and localization of viral infection in the pancreas of patients with type 1 diabetes using short fluorescently-labelled oligonucleotide probes

AU - Busse, Niels

AU - Paroni, Federico

AU - Richardson, Sarah J

AU - Laiho, Jutta E

AU - Oikarinen, Maarit

AU - Frisk, Gun

AU - Hyöty, Heikki

AU - de Koning, Eelco

AU - Morgan, Noel G

AU - Maedler, Kathrin

PY - 2017/2/21

Y1 - 2017/2/21

N2 - 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.

AB - 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.

KW - Diabetes Mellitus, Type 1

KW - Enterovirus

KW - Enterovirus Infections

KW - Fluorescent Dyes

KW - Humans

KW - Immunohistochemistry

KW - In Situ Hybridization, Fluorescence

KW - Oligonucleotide Probes

KW - Pancreas

KW - Polymerase Chain Reaction

KW - RNA, Viral

KW - Sensitivity and Specificity

KW - Journal Article

U2 - 10.18632/oncotarget.14896

DO - 10.18632/oncotarget.14896

M3 - Article

C2 - 28147344

SN - 1949-2553

VL - 8

SP - 12620

EP - 12636

JO - Oncotarget

JF - Oncotarget

IS - 8

ER -

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

Abstract

Keywords

Access to Document

Fingerprint

Cite this