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PCR detection of a hantavirus and Rift Valley fever virus using dried whole blood spotted on filter strips
Umeå University, Faculty of Medicine, Department of Clinical Microbiology, Infectious Diseases. Umeå University, Faculty of Medicine, Department of Clinical Microbiology, Virology. (Clas Ahlm)
Umeå University, Faculty of Medicine, Department of Clinical Microbiology, Virology.
Umeå University, Faculty of Medicine, Department of Clinical Microbiology, Virology.
Totalförsvarets forskningsinstitut, FOI, Umeå.
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(English)Manuscript (preprint) (Other academic)
Abstract [en]

Viral hemorrhagic fevers are serious and emerging infections among humans and animals worldwide. Presently, blood or serum samples are the main source for diagnostics. However, transportation of such samples from remote areas may be complicated and expensive. Previously, filter strips soaked with blood have been used for detection of antibodies for diagnostics and epidemiological studies of several infectious diseases.

In this study we evaluate if a similar approach could be applied for detection of viral RNA of Rift Valley Fever virus or Hantavirus (Puumala).

We have used whole blood spiked with known amounts of viruses. In addition, clinical samples from patients with acute hemorrhagic fever with renal syndrome have been analysed. The samples were collected on filter strips and dried before RNA was extracted at different time-points. For Puumala, the sensitivity was acceptable, although the absolute levels of viral RNA were found to be considerable lower when using filter strips. The viral RNA could be detected and analysed after 2-3 weeks storage of the dried filter strips. In contrast, for RVFV, no or very low copy numbers of viral RNA were detected. Still, RVFV filter strips contained viable virus particles up to 48 h of storage.

In conclusion, the use of dried whole blood samples spotted on filter strips for the detection of viral RNA seems to be a reliable and simple procedure for Hantaviruses. This procedure could be useful in field studies, especially in remote areas or low-income countries where transportation and storage of biological samples might be complicated. However, the result for RVFV was unexpected and further studies are needed to improve this technique.

Keyword [en]
Rift Valley fever, hantavirus, filter strips
Research subject
Medical Virology
URN: urn:nbn:se:umu:diva-30673OAI: diva2:285526
Ej publicerat. Not submitted.Available from: 2010-01-12 Created: 2010-01-12 Last updated: 2011-10-14Bibliographically 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. 64 p.
Umeå University medical dissertations, ISSN 0346-6612 ; 1323
Rift Valley Fever, vaccines, diagnostics
National Category
Medical and Health Sciences Microbiology in the medical area
Research subject
Medical Virology; Infectious Diseases
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)
Available from: 2010-01-18 Created: 2010-01-12 Last updated: 2010-01-18Bibliographically approved

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Näslund, JonasDrobni, PeterKerner, AlexanderEvander, MagnusAhlm, Clas
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