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Kinetics of Rift Valley fever virus in experimentally infected mice using quantitative real-time RT-PCR
Umeå University, Faculty of Medicine, Department of Clinical Microbiology, Virology. Umeå University, Faculty of Medicine, Department of Clinical Microbiology, Infectious Diseases.
Umeå University, Faculty of Medicine, Department of Clinical Microbiology, Infectious Diseases. Umeå University, Faculty of Medicine, Department of Clinical Microbiology, Virology.
Umeå University, Faculty of Medicine, Department of Clinical Microbiology, Virology.
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2008 (English)In: Journal of Virological Methods, ISSN 0166-0934, E-ISSN 1879-0984, Vol. 151, no 2, p. 277-282Article in journal (Refereed) Published
Abstract [en]

Rift Valley Fever (RVF) is an important viral zoonosis in Africa affecting animals and humans. Since no protective vaccines or effective treatments are available for human use, accurate and reliable diagnostic methods are essential for surveillance of the disease in order to implement adequate public health actions. To study the kinetics of the RVF Virus (RVFV) infection, a SYBR Green-based quantitative real-time RT-PCR assay was developed. By using primers targeting the S-segment of RVFV, the detection limit of this assay was estimated to 30 RNA templates. Blood and organs of experimentally infected mice were sampled at different time points and RVFV RNA was quantified. High amounts of RVFV RNA were found in blood, brain, and liver samples shortly after infection with a 1-4 days post infection window for viral RNA detection. Mice developed symptoms after the appearance of serum antibodies, indicating that the host response plays an important role in the outcome of the disease. The RVFV quantitative RT-PCR proved to be a valuable diagnostic tool during the first days of infection, before detectable antibody levels and visual symptoms of RVF were observed.

Place, publisher, year, edition, pages
2008. Vol. 151, no 2, p. 277-282
Identifiers
URN: urn:nbn:se:umu:diva-21109DOI: 10.1016/j.jviromet.2008.04.007PubMedID: 18514921Scopus ID: 2-s2.0-46949107190OAI: oai:DiVA.org:umu-21109DiVA, id: diva2:210608
Available from: 2009-04-02 Created: 2009-04-02 Last updated: 2023-03-24Bibliographically approved
In thesis
1. Rift Valley fever: development of diagnostics and vaccines
Open this publication in new window or tab >>Rift Valley fever: development of diagnostics and vaccines
2010 (English)Doctoral thesis, comprehensive summary (Other academic)
Abstract [en]

Rift Valley Fever virus (RVFV) causes an infection with severe impact on animal and human health. The disease is endemic throughout almost the entire African continent and large regions of the Arabian Peninsula. During epidemics, high mortality is observed in animals, especially among cattle, goats, and sheep. In humans, the symptoms vary from a benign influenza-like disease to a life-threatening hemorrhagic fever. Due to the devastating effect on communities in endemic regions and the possibility of further spread of this virus, there is an imperative need to improve and develop control measurements against this emerging disease. Therefore, this thesis focuses on diagnostics and vaccines against RVFV.

RVFV infection kinetics was studied in a mouse model system by detection and quantification of viral genomes, using a developed quantitative real-time PCR (QRT-PCR) method. This novel QRT-PCR method proved to be reliable and serves as a supplement to standard diagnostics, direct virus isolation and serological methods. High levels of viral RNA were found in blood and liver samples from experimentally infected mice during the first days post infection. Thereafter the levels declined rapidly and dropped below detection limit approximately seven days post infection. The QRT-PCR technique was also used in a study aimed to improve diagnosis of RVFV from field samples collected on filter strips.

Today, the available RVFV vaccines are only approved for animal use and these vaccines have several shortcomings. Since RVFV is a highly pathogenic organism requiring bio-safety level 3 laboratories, two different none-replicating vaccine approaches have been applied and evaluated using a mouse model. A DNA based vaccine, administered via gene-gun, and the use of virus-like particles (VLP), by the intra-peritoneal route. RVFV specific and neutralising antibodies were raised with both vaccine approaches. However, VLP vaccination against Rift valley Fever proved to be more promising as a future vaccine, since higher titres of neutralising antibodies and improved survival rate were found upon a lethal RVFV challenge in mice.

In conclusion, a sensitive and specific method for quantifying RVFV infection and a promising vaccine candidate against RVFV were developed.

Place, publisher, year, edition, pages
Umeå: Umeå university, 2010. p. 64
Series
Umeå University medical dissertations, ISSN 0346-6612 ; 1323
Keywords
Rift Valley Fever, vaccines, diagnostics
National Category
Medical and Health Sciences Microbiology in the medical area
Research subject
Medical Virology; Infectious Diseases
Identifiers
urn:nbn:se:umu:diva-30676 (URN)978-91-7264-883-8 (ISBN)
Public defence
2010-02-05, Betula, byggnad 6M, 901 85 umeå universitet, NUS, 09:00 (English)
Opponent
Supervisors
Available from: 2010-01-18 Created: 2010-01-12 Last updated: 2018-06-08Bibliographically approved

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Näslund, JonasEvander, MagnusAhlm, Clas

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