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Quantitative real-time PCR assay for detection of human polyomavirus infection.
Uppsala University, Medicinska vetenskapsområdet, Faculty of Medicine, Department of Medical Sciences.
Uppsala University, Medicinska vetenskapsområdet, Faculty of Medicine, Department of Medical Sciences. (Klinisk Virologi. Blomberg)
2006 (English)In: J Virol Methods, ISSN 0166-0934, Vol. 135, no 2, 207-13 p.Article in journal (Refereed) Published
Abstract [en]

The overall aim was to study factors that affect behaviour related to CVD (cardiovascular diseases). Study I tested whether gender, education and so-cioeconomic status correlated to knowledge about risk factors, and Study II studied knowledge and risk behaviour from a national perspective (Sweden versus Poland). Furthermore, Study III examined whether obese people dif-fered from people of normal weight regarding knowledge about risk factors, and Study IV examined whether risk behaviour is affected by personal ex-perience of illness and family history of CVD.

The studies are population-based with cross-sectional design. Data were obtained by questionnaires and by screening results of risk factors related to CVD. The studies were carried out among 50-year old men and women in Västmanland, Sweden (n=1011) and in Wroclaw, Poland (n=1043).

The results show that women are more knowledgeable than men about the risk factors for CVD, and that low education is associated with insufficient knowledge about CVD (Study I). The discrepancy between knowledge and behaviour was greater among the Poles than it was among the Swedes (Study II). Obese individuals did not differ significantly from individuals with a normal weight regarding knowledge of cardiovascular risk factors when education was controlled for (Study III). Individuals with a personal experience of illness may be more inclined to change smoking behaviour than the average person (Study IV).

In conclusion, knowledge about risk factors for CVD varies with education, gender and, to a certain degree, nationality. However, knowledge does not only consist of the conditions of behaviour change. The results in the thesis substantiate theories suggesting that change in risk behaviour is a process over time. Predictors of risk behaviours on the individual level as well as national level are of importance, and needs to be considered in the every day practice of health care professionals.

Place, publisher, year, edition, pages
2006. Vol. 135, no 2, 207-13 p.
Keyword [en]
BK Virus/isolation & purification, Base Sequence, Cerebrospinal Fluid/virology, DNA Primers, DNA; Viral/analysis, Humans, JC Virus/isolation & purification, Molecular Sequence Data, Polymerase Chain Reaction/*methods, Polyomavirus Infections/*diagnosis, Sensitivity and Specificity, Simian virus 40/isolation & purification
URN: urn:nbn:se:uu:diva-11884PubMedID: 16677718OAI: oai:DiVA.org:uu-11884DiVA: diva2:39653
Available from: 2008-06-27 Created: 2008-06-27 Last updated: 2011-01-11
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.
Digital Comprehensive Summaries of Uppsala Dissertations from the Faculty of Medicine, ISSN 1651-6206 ; 202
Microbiology, Real-time PCR, broadly targeted PCR, degenerated PCR, rTth DNA polymerase, thermolabile UNG, QPIA, IgM, Mikrobiologi
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)
Available from: 2006-11-21 Created: 2006-11-21 Last updated: 2009-05-08Bibliographically approved

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