Development of a real-time RT-PCR assay for improved detection of Borna disease virus.
Abstract: Borna disease virus (BDV) is a non-segmented, negative-stranded RNA virus, which infects cells of the central nervous system (CNS) in many different species. BDV is the causative agent of the neurological disorders in horses and sheep termed classical Borna disease (BD), as well as staggering disease in cats. At present, the diagnosis staggering disease or feline BD is made by histopathology or immunohistochemistry of the CNS. In order to obtain a better clinical diagnostic tool, a duplex real-time RT-PCR assay (rRT-PCR) was developed. TaqMan probes and primers specific for the BDV P and BDV L genes were designed by aligning the sequences of known BDV strains. After optimisation, the sensitivity and specificity of the rRT-PCR were established. The detection limit was set to 10-100 viral genomic copies per reaction and the assay detects the BDV strains V and He/80, as well as the most divergent BDV strain known so far, No/98. Furthermore, the system detected feline BDV variants in five naturally infected cats and a feline isolate used in experimental infection of cats. This rRT-PCR assay will be a powerful tool in further studies of BDV, including epidemiological screening and diagnosis.
Publication Date: 2007-03-21 PubMed ID: 17376545DOI: 10.1016/j.jviromet.2007.01.034Google Scholar: Lookup
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- Journal Article
- Research Support
- Non-U.S. Gov't
Summary
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The research article discusses the development of a real-time RT-PCR assay for superior detection of Borna Disease Virus (BDV), a virus that affects the central nervous system in various species like horses, sheep, and cats.
Background of Borna Disease Virus (BDV)
- BDV is categorized as a non-segmented, negative-stranded RNA virus.
- It primarily affects the cells of the central nervous system (CNS) across various species, including horses, sheep, and cats.
- In horses and sheep, BDV infection results in neurological disorders collectively known as classical Borna disease (BD).
- In cats, the disease caused by the BDV infection is referred to as ‘staggering disease’ or feline BD.
Current Diagnostic Methods
- At present, methods such as histopathology or immunohistochemistry of the CNS are used for diagnosing BDV infection in cats or the staggering disease.
- However, these methods may not be very advanced or accurate.
Development of a Duplex Real-time RT-PCR Assay
- To enhance the clinical diagnostic process, the research team developed a duplex real-time RT-PCR assay (rRT-PCR).
- This was achieved by designing specific TaqMan probes and primers for the BDV P and BDV L genes. The design was based on aligning sequences of known BDV strains.
Optimisation, Sensitivity, and Specificity of the rRT-PCR Assay
- Post optimization, the sensitivity and specificity of the rRT-PCR assay were established.
- The detection limit was set to between 10-100 viral genomic copies per reaction.
- The rRT-PCR assay is proficient in detecting various BDV strains including the most diverse strain known as No/98, along with BDV strains V and He/80.
Detection of Feline BDV Variants
- The developed system was successful in detecting feline BDV variants in five naturally infected cats.
- Additionally, it was used to detect a feline isolate obtained from an experimental infection of cats.
Implications of the Developed rRT-PCR Assay
- With its advanced detection capabilities, this rRT-PCR assay can significantly contribute to deeper studies of the BDV, aiding in aspects like epidemiological screening and diagnosis.
Cite This Article
APA
Wensman JJ, Thorén P, Hakhverdyan M, Belák S, Berg M.
(2007).
Development of a real-time RT-PCR assay for improved detection of Borna disease virus.
J Virol Methods, 143(1), 1-10.
https://doi.org/10.1016/j.jviromet.2007.01.034 Publication
Researcher Affiliations
- Joint Research and Development Division in Virology of National Veterinary Institute (SVA), Swedish University of Agricultural Sciences (SLU), Uppsala, Sweden. Jonas.Wensman@bvf.slu.se
MeSH Terms
- Animals
- Base Sequence
- Borna Disease / diagnosis
- Borna Disease / virology
- Borna disease virus / genetics
- Borna disease virus / isolation & purification
- Borna disease virus / pathogenicity
- Cat Diseases / diagnosis
- Cat Diseases / psychology
- Cats
- Genes, Viral
- Molecular Sequence Data
- Reverse Transcriptase Polymerase Chain Reaction / methods
- Sensitivity and Specificity
Citations
This article has been cited 12 times.- Ashwell M, Freire M, O'Nan AT, Benito J, Hash J, McCulloch RS, Lascelles BDX. Characterization of gene expression in naturally occurring feline degenerative joint disease-associated pain. Vet J 2019 Jan;243:42-47.
- Pinto MC, Rondahl V, Berg M, Ågren E, Carvalheira J, Thompson G, Johansson Wensman J. Detection and phylogenetic analysis of parrot bornavirus 4 identified from a Swedish Blue-winged macaw (Primolius maracana) with unusual nonsuppurative myositis. Infect Ecol Epidemiol 2019;9(1):1547097.
- Formisano S, Hornig M, Yaddanapudi K, Vasishtha M, Parsons LH, Briese T, Lipkin WI, Williams BL. Central Nervous System Infection with Borna Disease Virus Causes Kynurenine Pathway Dysregulation and Neurotoxic Quinolinic Acid Production. J Virol 2017 Jul 15;91(14).
- Björnsdóttir S, Agustsdóttir E, Blomström AL, Oström IL, Berndtsson LT, Svansson V, Wensman JJ. Serological markers of Bornavirus infection found in horses in Iceland. Acta Vet Scand 2013 Nov 1;55(1):77.
- Zhang L, Xu MM, Zeng L, Liu S, Liu X, Wang X, Li D, Huang RZ, Zhao LB, Zhan QL, Zhu D, Zhang YY, Xu P, Xie P. Evidence for Borna disease virus infection in neuropsychiatric patients in three western China provinces. Eur J Clin Microbiol Infect Dis 2014 Apr;33(4):621-7.
- Kinnunen PM, Inkeroinen H, Ilander M, Kallio ER, Heikkilä HP, Koskela E, Mappes T, Palva A, Vaheri A, Kipar A, Vapalahti O. Intracerebral Borna disease virus infection of bank voles leading to peripheral spread and reverse transcription of viral RNA. PLoS One 2011;6(8):e23622.
- Song JW, Na KS, Tae SH, Kim YK. Borna disease virus antibody and RNA from peripheral blood mononuclear cells of race horses and jockeys in Korea. Psychiatry Investig 2011 Mar;8(1):58-60.
- Belyi VA, Levine AJ, Skalka AM. Unexpected inheritance: multiple integrations of ancient bornavirus and ebolavirus/marburgvirus sequences in vertebrate genomes. PLoS Pathog 2010 Jul 29;6(7):e1001030.
- Na KS, Tae SH, Song JW, Kim YK. Failure to detect borna disease virus antibody and RNA from peripheral blood mononuclear cells of psychiatric patients. Psychiatry Investig 2009 Dec;6(4):306-12.
- Thilén E, Rubbenstroth D, Tengstrand S, Pfaff F, Wensman JJ, Ley C. Evidence of rustrela virus-associated feline staggering disease in Sweden since the 1970s. Acta Vet Scand 2024 Nov 23;66(1):59.
- Wensman JJ, Jäderlund KH, Gustavsson MH, Hansson-Hamlin H, Karlstam E, Lilliehöök I, Oström IL, Belák S, Berg M, Holst BS. Markers of Borna disease virus infection in cats with staggering disease. J Feline Med Surg 2012 Aug;14(8):573-82.
- De Risio L, Brown R, Tennant B, Sparkes A, Matiasek L, de Stefani A, Weissenböck H, Matiasek K. Slowly progressive lymphohistiocytic meningoencephalomyelitis in 21 adult cats presenting with peculiar neurological signs. J Feline Med Surg 2012 Apr;14(4):250-6.
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