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3 : Laboratory Diagnosis

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Abstract:

Laboratory diagnosis of Borna disease virus (BDV) has increasingly become a crucial issue in BDV research during the past 15 years. The methods of intra vitam diagnosis of BDV infection in animals that display neurological symptoms typical for Borna disease (BD) have gained general acceptance. Serology is still the most commonly used intra vitam detection method. Serological assays include indirect immunofluorescence assay (IFA), Western blot assay (WBA) and enzyme-linked immunosorbent assay (ELISA). To directly demonstrate infection with BDV, several methods to detect BDV antigens are in use. Virus antigen detection includes Immunohistochemistry (IHC), WBA and fluorescence activated cell sorting (FACS). Since mRNAs coding for the viral nucleoprotein (p40; N) and phosphoprotein (p24; P) are the most abundant viral transcripts in BDV-infected tissues and cells, almost all RNA detection studies have aimed to detect BDV Nor P-specific sequences. Virus RNA detection includes Northern Hybridization, RT-PCR and RT-Nested PCR. For analysis of virus infectivity, both in vitro and in vivo systems have been established. The application of PCR techniques to BDV research yielded a wealth of information regarding sequence variability and diagnosis of BDV infection. Using RT-PCR to detect virus RNA in brains of animals with BD, the majority of studies could detect RNA in all cases analyzed. Sequencing of the RT-PCR or RT–nested-PCR products might help to define whether PCR products were amplified from laboratory BDV plasmids or contaminating BDV RNA or whether they represent new BDV strains.

Citation: Sauder C, Yamaguchi K, Mizutani T. 2002. Laboratory Diagnosis, p 45-85. In Carbone K (ed), Borna Disease Virus and Its Role in Neurobehavioral Diseases. ASM Press, Washington, DC. doi: 10.1128/9781555817909.ch3

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Figure 1

IFA to detect BDV-reactive antibodies in serum from a diseased horse (A and B) and a psychiatric patient (C and D), using C6 cells (A and C) and persistently BDV-infected C6 cells (B and D). Both sera contain BDV-reactive antibodies as judged by the punctate staining pattern in nuclei of infected cells. (Courtesy of Christian Billich.)

Citation: Sauder C, Yamaguchi K, Mizutani T. 2002. Laboratory Diagnosis, p 45-85. In Carbone K (ed), Borna Disease Virus and Its Role in Neurobehavioral Diseases. ASM Press, Washington, DC. doi: 10.1128/9781555817909.ch3
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