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Experience from the development of a diagnostic single tube real-time PCR for human caliciviruses, Norovirus genogroups I and II.
Uppsala University, Medicinska vetenskapsområdet, Faculty of Medicine, Department of Medical Sciences. (Klinisk Virologi. Blomberg)
Uppsala University, Medicinska vetenskapsområdet, Faculty of Medicine, Department of Medical Sciences. (Klinisk Virologi. Blomberg)
2006 (English)In: J Virol Methods, ISSN 0166-0934, Vol. 132, no 1-2, 69-76 p.Article in journal (Refereed) Published
Place, publisher, year, edition, pages
2006. Vol. 132, no 1-2, 69-76 p.
Keyword [en]
Adult, Caliciviridae Infections/*diagnosis/virology, Child, DNA Primers, Feces/virology, Gastroenteritis/virology, Humans, Microscopy; Electron, Norovirus/*classification/genetics/*isolation & purification, RNA; Viral/genetics, Reverse Transcriptase Polymerase Chain Reaction/*methods
URN: urn:nbn:se:uu:diva-11885PubMedID: 16289337OAI: oai:DiVA.org:uu-11885DiVA: diva2:39654
Available from: 2008-06-25 Created: 2008-06-25 Last updated: 2011-01-11
In thesis
1. Molecular Diagnosis of Common Viral Infectious Diseases Based on Real-Time PCR
Open this publication in new window or tab >>Molecular Diagnosis of Common Viral Infectious Diseases Based on Real-Time PCR
2006 (English)Doctoral thesis, comprehensive summary (Other academic)
Abstract [en]

Molecular biology has become an integral part of the diagnosis of infectious diseases. Recently, quantitative real-time PCR (QPCR) methods (often in the form of so-called TaqMan® systems) have been developed for the diagnosis of a wide range of infectious diseases; these techniques found valuable clinical application in the diagnosis and evaluation of progress and therapeutic success of viral diseases. The use of QPCR as a tool for diagnostic virological and viral research laboratories has greatly increased in recent years. It often replaces conventional PCR and amplicon detection systems which are more complex and laborious, with a higher risk of amplicon carry-over contamination.

The new QPCR methods presented here utilize broadly targeted primers and probes for rational and sensitive detection and quantification of variable RNA viruses. They take advantage of the dual properties, both RNA and DNA dependent DNA polymerase activities, of the rTth thermostable polymerase, and thermolabile UNG with dUTP to protect against inadvertent contamination of samples with amplimers.

In paper one, a novel QPCR approach to detect and quantify human enteroviral (EV) RNA in patients with neurological disorders such as aseptic meningitis is presented. In the second paper, the development of a novel serological technique, quantitative PCR enhanced immunoassay (QPIA), for serodiagnosis of EV infection, is described. In paper three the subject is the development of a touch-down QPCR (TD-QPCR) for detection and preliminary genogrouping of norovirus (NV), a group of Caliciviruses. In paper four a rational, broadly targeted, system for detection of diverse influenza viruses, yet being able to discriminate between influenza A, B and C, is designed and evaluated. In the last paper, another rational broadly targeted system, for detection of corona viruses in humans and animals, is described.

The technologies described in this collection of papers have common features. They are a platform for further development of diagnostic tools for screening and detection of viruses in known viral diseases, maybe also for discovering new viruses.

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 ; 168
Molecular biology, Microbiology, Virology, rTth DNA polymerase, UNG, QPCR, RNA virus, broadly targeted PCR, Molekylärbiologi
urn:nbn:se:uu:diva-7118 (URN)91-554-6639-7 (ISBN)
Public defence
2006-10-06, , Medical Microbiology, Dag Hammar Skjölds vag. 17, Uppsala, 09:15 (English)
Available from: 2006-09-18 Created: 2006-09-18 Last updated: 2011-02-28Bibliographically approved

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