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Recent enterovirus infection in type 1 diabetes: evidence with a novel IgM method
Uppsala University, Disciplinary Domain of Medicine and Pharmacy, Faculty of Medicine, Department of Medical Sciences.
Uppsala University, Disciplinary Domain of Medicine and Pharmacy, Faculty of Medicine, Department of Women's and Children's Health.
Uppsala University, Disciplinary Domain of Medicine and Pharmacy, Faculty of Medicine, Department of Women's and Children's Health. (Barnendokrinologisk forskning/Gustafsson)
Uppsala University, Disciplinary Domain of Medicine and Pharmacy, Faculty of Medicine, Department of Women's and Children's Health.
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2007 (English)In: Journal of Medical Virology, ISSN 0146-6615, E-ISSN 1096-9071, Vol. 79, no 12, 1861-1867 p.Article in journal (Refereed) Published
Abstract [en]

Enterovirus (EV) infection has been associated with Type 1 (T1D) diabetes and on a few occasions virus could be isolated at onset of the disease. Using two such isolates as antigens in a quantitative PCR enhanced immunoassay (T1D-EV-QPIA) we have measured IgM antibodies against such potentially diabetogenic viruses in serum from 33 newly diagnosed T1D children, 24 siblings, and 27 healthy children. Sera were also analysed with regard to autoantibodies against GAD65, the cytokine TNF-alpha and the chemokine IP-10. EV-RNA detection was performed on peripheral blood mononuclear cells (PBMC). IgM antibodies against this "new" EV antigen were more frequent in serum from T1D children than in serum from siblings and/or controls (P < 0.001). EV-RNA was detected more frequently in PBMC from T1D children than in healthy control children (P < 0.001) and also compared to the siblings (P < 0.003). The cytokine TNF-alpha was less frequently detected in serum from the T1D children compared with serum from siblings and/controls (P < 0.001). A positive correlation was found between the results obtained with the T1D-EV-QPIA and the EV-PCR (P < 0.001). These findings are in line with earlier findings of an increased frequency of enteroviral infections in newly diagnosed T1D patients. In addition, we found that T1D children at onset of the disease had lower frequencies of the chemokine TNF-alpha in their serum than age- and sex-matched controls had, suggesting an impaired immune response.

Place, publisher, year, edition, pages
2007. Vol. 79, no 12, 1861-1867 p.
Keyword [en]
enterovirus, type 1 diabetes, quantitative PCR enhanced immunoassay, IgM antibodies, autoantibodies against GAD65, TNF-
National Category
Medical and Health Sciences
Identifiers
URN: urn:nbn:se:uu:diva-11874DOI: 10.1002/jmv.21008ISI: 000250319100011PubMedID: 17935175OAI: oai:DiVA.org:uu-11874DiVA: diva2:39643
Available from: 2008-06-11 Created: 2008-06-11 Last updated: 2017-12-11Bibliographically approved
In thesis
1. Development of Real-Time PCR Based Methods for Detection of Viruses and Virus Antibodies
Open this publication in new window or tab >>Development of Real-Time PCR Based Methods for Detection of Viruses and Virus Antibodies
2006 (English)Doctoral thesis, comprehensive summary (Other academic)
Abstract [en]

Quantitative real-time PCR (QPCR) technology has been very useful for diagnosis of viral diseases. QPCR has recently reached a level of sensitivity, simplicity, and reproducibility which allows a large number of samples to be screened rapidly, make it a suitable tool for the clinical virology diagnostics.

In this thesis, broadly targeted and degenerated quantitative QPCR assays were used. A somewhat novel single-tube real-time reverse transcription-polymerase chain reaction (QRT-PCR), with takes advantage of ability of rTth DNA polymerase to reverse transcribe RNA in the presence of Mn2+ at elevated temperatures and includes protection against amplimer contamination by using thermolabile UNG, was developed. A new technique for diagnostic of recent viral infection by detection of viral immunoglobulin M (IgM) was also developed.

In the first paper, a sensitive single-tube QRT-PCR for detection of enteroviral RNA in patients with aseptic meningitis was presented. In the second paper, a single-serum-dilution real-time PCR-based PIA (PCR-enhanced immunoassay), called quantitative PIA (QPIA), to detect enterovirus IgM for diagnosis of EV infection in patients with aseptic meningitis, was also developed. In the third paper, a broadly targeted, simple, single tube degenerated quantitative QPCR technique for detection of JCV, BKV and SV40 DNA was developed. A conserved region of the VP2 gene of JCV, BKV and SV40 was targeted. A false positive result due to contamination with commonly used SV40 T-antigen plasmids was therefore avoided. In manuscript four, the QPIA assay provide a rational strategy for detection of EV IgM, allows the use of viral antigens isolate from newly diagnosed Type 1 diabetes patients (T1D-EV-QPIA) to measured IgM against diabetogenic viruses in serum from newly diagnosed T1D children, siblings, and healthy children.

To conclude, novel broadly targeted real-time PCR methods for diagnosis of entero- and polyoma viral infections were developed.

Place, publisher, year, edition, pages
Uppsala: Acta Universitatis Upsaliensis, 2006. 60 p.
Series
Digital Comprehensive Summaries of Uppsala Dissertations from the Faculty of Medicine, ISSN 1651-6206 ; 202
Keyword
Microbiology, Real-time PCR, broadly targeted PCR, degenerated PCR, rTth DNA polymerase, thermolabile UNG, QPIA, IgM, Mikrobiologi
Identifiers
urn:nbn:se:uu:diva-7320 (URN)91-554-6725-3 (ISBN)
Public defence
2006-12-12, Hörsalen, ing, D1, Dag Hammarskjölds Väg 17, Akademiska Sjukhuset 751 85, UPPSALA, 13:15 (English)
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Available from: 2006-11-21 Created: 2006-11-21 Last updated: 2009-05-08Bibliographically approved

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Frisk, GunYin, HongTuvemo, TorstenBlomberg, Jonas

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