Change search
ReferencesLink to record
Permanent link

Direct link
Microarray technology for identification and distinction of hantaviruses.
Umeå University, Faculty of Medicine, Clinical Microbiology, Virology.
Umeå University, Faculty of Medicine, Clinical Microbiology, Virology.
Show others and affiliations
2004 (English)In: Journal of Medical Virology, ISSN 0146-6615, Vol. 72, no 4, 646-655 p.Article in journal (Refereed) Published
Abstract [en]

DNA microarrays combine high-precision technology with advanced molecular biology to achieve high-throughput screening of DNA fragments. In this study, we investigated the potential of the cDNA microarray technique to identify and discriminate PCR derived amplicons from genetically highly similar viruses. The wide range of sequence variation among hantaviruses makes them suitable as a model for this purpose. The hantaviruses, carried by rodents, cause several hundred thousand cases of severe human disease every year in many parts of the world. A hantavirus-specific microarray, including DNA fragments from 12 viral isolates of six different hantaviruses, was designed. The S and M genome segments were represented by 500-nucleotide overlapping and 250-nucleotide non-overlapping fragments. A considerable ability to distinguish between different hantaviruses was demonstrated using a novel analysis method. Even different isolates of a single virus, were identified correctly despite 90% sequence similarity. The distinction ability was accompanied by a tolerance for smaller sequence differences, which makes the microarray suitable for testing samples containing unknown viruses. Viral genetic material found in samples from the lungs of bank voles caught in the wild was identified precisely, which demonstrated further the potential for this technology.

Place, publisher, year, edition, pages
2004. Vol. 72, no 4, 646-655 p.
Keyword [en]
Animals, Arvicolinae/*virology, Genes; Viral, Genome; Viral, Hantaan virus/classification/genetics, Hantavirus/*classification/*genetics/isolation & purification, Hantavirus Infections/veterinary/virology, Lung/virology, Molecular Probe Techniques, Nucleic Acid Hybridization/*methods, Oligonucleotide Array Sequence Analysis/*methods, Polymerase Chain Reaction, Puumala virus/classification/genetics, RNA; Viral/chemistry/isolation & purification, Rodent Diseases/virology, Seoul virus/classification/genetics, Sin Nombre virus/classification/genetics, Transcription; Genetic, Variation (Genetics)
URN: urn:nbn:se:umu:diva-20713DOI: 10.1002/jmv.20041PubMedID: 14981768OAI: diva2:209388
Available from: 2009-03-24 Created: 2009-03-24 Last updated: 2009-11-16
In thesis
1. Implications of Local Puumala Hantavirus Genetics and Epidemiology for Diagnostics and Vaccine Development
Open this publication in new window or tab >>Implications of Local Puumala Hantavirus Genetics and Epidemiology for Diagnostics and Vaccine Development
2005 (English)Doctoral thesis, comprehensive summary (Other academic)
Abstract [en]

Puumala viruses, a member of the Hantavirus genus in the Bunyaviridae family, are enveloped by a lipid bilayer and possesses a tripartite single stranded RNA genome with negative polarity. The hantaviruses encode four proteins: a nucleocapsid protein (N), two membrane spanning glycoproteins (GN and GC) and a RNA dependent RNA polymerase (RdRp). Hantaviruses cause two forms of diseases, hemorrhagic fever with renal syndrome (HFRS) in Europe and Asia, and hantavirus pulmonary syndrome (HPS) in the Americas. The hantaviruses are mainly rodent borne, and humans are mostly infected by inhalation of aerosolized rodent secrete. Human Puumala virus infection results in nephropathia epidemica (NE), a mild haemorrhagic disease.

It is of importance to have a good understanding of the epidemiology and genetics of these viruses for the development of new diagnostic methods and for future vaccine development.

In this thesis we determined the complete viral genome sequence and characterized the structural proteins based on studies of expression and glycosylation patterns, for a unique human virus isolate; performed a genomic analysis of local Puumala viruses and their individual rodent host, Clethrionomys glareolus, from six different locations was performed. It was seen that the virus genetic variation between different locations could be stable over relatively large distances while there could be large variation over a short distance. For the bank voles no such variation could be seen; developed and evaluated Genetic vaccines, based on PCR-generated linear DNA. We showed that it was important to protect these fragments against nuclease degradation at that attachment of a nuclear localization signal peptide further improved the immune response. We also designed, fabricated and evaluated a 2000 probe cDNA-microarray for identification and differentiation of hantaviruses. The chips was based on 12 different strains of six hantaviruses and could differentiate between both different hantaviruses and strains within one hantavirus serotype.

Place, publisher, year, edition, pages
Umeå: Klinisk mikrobiologi, 2005. 97 p.
Umeå University medical dissertations, ISSN 0346-6612 ; 964
Communicable diseases, hantavirus, Puumala virus, nephropathia epidemica, genome sequence, glycosylation, linear DNA vaccine, microarray, identification, genetic variability, Infektionssjukdomar
National Category
Infectious Medicine
Research subject
Medical Virology
urn:nbn:se:umu:diva-532 (URN)91-7305-878-5 (ISBN)
Public defence
2005-06-03, A05, 6A, Umeå Universitet, Umeå, 09:00 (English)
Available from: 2005-05-09 Created: 2005-05-09 Last updated: 2009-11-16Bibliographically approved

Open Access in DiVA

No full text

Other links

Publisher's full textPubMed
By organisation
In the same journal
Journal of Medical Virology

Search outside of DiVA

GoogleGoogle Scholar
The number of downloads is the sum of all downloads of full texts. It may include eg previous versions that are now no longer available

Altmetric score

Total: 21 hits
ReferencesLink to record
Permanent link

Direct link