Health care professionals at risk of infection with Borna disease virus - evidence from a large hospital in China (Chongqing).

BACKGROUND: Human Borna disease virus (BDV) infections have recently been reported in China. BDV causes cognitive and behavioural disturbances in animals. The impact on human mental disorders is subject to debate, but previous studies worldwide have found neuropsychiatric patients more frequently infected than healthy controls. A few isolates were recovered from severely depressed patients, but contagiousness of BDV strain remains unknown. METHOD: We addressed the risk of infection in health care settings at the first affiliated hospital of Chongqing Medical University (CQMU), located in downtown Chongqing, a megacity in Southwest China. Between February 2012 and March 2013, we enrolled 1529 participants, of whom 534 were outpatients with major depressive disorder (MDD), 615 were hospital personnel, and 380 were healthy controls who underwent a health check. Infection was determined through BDV-specific circulating immune complexes (CIC), RNA, and selective antibodies (blood). RESULTS: One-fifth of the hospital staff (21.8%) were found to be infected (CIC positive), with the highest prevalence among psychiatry and oncology personnel, which is twice as many as were detected in the healthy control group (11.1%), and exceeds the prevalence detected in MDD patients (18.2%). CONCLUSION: BDV circulates unnoticed in hospital settings in China, putting medical staff at risk and warranting clarification of infection modes and introduction of prevention measures.

Borna disease virus (BDV) infection in psychiatric patients and healthy controls in Iran.

BACKGROUND: Borna disease virus (BDV) is an evolutionary old RNA virus, which infects brain and blood cells of humans, their primate ancestors, and other mammals. Human infection has been correlated to mood disorders and schizophrenia, but the impact of BDV on mental-health still remains controversial due to poor methodological and cross-national comparability. METHOD: This first report from the Middle East aimed to determine BDV infection prevalence in Iranian acute psychiatric disorder patients and healthy controls through circulating immune complexes (CIC), antibodies (Ab) and antigen (pAg) in blood plasma using a standardized triple enzyme immune assay (EIA). Samples of 314 subjects (114 psychiatric cases, 69 blood donors, and 131 healthy controls) were assayed and data analyzed quantitatively and qualitatively. RESULTS: CICs revealed a BDV prevalence of one third (29.5%) in healthy Iranian controls (27.5% controls; 33.3% blood donors). In psychiatric patients CIC prevalence was higher than in controls (40.4%) and significantly correlating with bipolar patients exhibiting overt clinical symptoms (p = 0.005, OR = 1.65). CIC values were significantly elevated in bipolar (p = 0.001) and major depressive disorder (p = 0.029) patients as compared to controls, and in females compared to males (p = 0.031). CONCLUSION: This study supports a similarly high prevalence of subclinical human BDV infections in Iran as reported for central Europe, and provides again an indication for the correlation of BDV infection and mood disorders. Further studies should address the morbidity risk for healthy carriers and those with elevated CIC levels, along with gender disparities.

Evidence for natural Borna disease virus infection in healthy domestic animals in three areas of western China.

Borna disease virus (BDV) is a non-cytolytic, neurotropic RNA virus that can infect many vertebrate species, including humans. To date, BDV infection has been reported in a range of animal species across a broad global geographic distribution. However, a systematic epidemiological survey of BDV infection in domesticated animals in China has yet to be performed. In current study, BDV RNA and antibodies in 2353 blood samples from apparently healthy animals of eight species (horse, donkey, dog, pig, rabbit, cattle, goat, sheep) from three areas in western China (Xinjiang province, Chongqing municipality, and Ningxia province) were assayed using reverse transcription qPCR (RT-qPCR) and ELISA assay. Brain tissue samples from a portion of the BDV RNA- and/or antibody-positive animals were subjected to RT-qPCR and western blotting. As a result, varying prevalence of BDV antibodies and/or RNA was demonstrated in various animal species from three areas, ranging from 4.4 % to 20.0 %. Detection of BDV RNA and/or antibodies in Chongqing pigs (9.2 %) provided the first known evidence of BDV infection in this species. Not all brain tissue samples from animals whose blood was BDV RNA and/or antibody positive contained BDV RNA and protein. This study provides evidence that BDV infection among healthy domestic animal species is more widespread in western China than previously believed.

Antibodies against viral nucleo-, phospho-, and X protein contribute to serological diagnosis of fatal Borna disease virus 1 infections.

Borna disease virus 1 (BoDV-1) causes rare but often fatal encephalitis in humans. Late diagnosis prohibits an experimental therapeutic approach. Here, we report a recent case of fatal BoDV-1 infection diagnosed on day 12 after hospitalization by detection of BoDV-1 RNA in the cerebrospinal fluid. In a retrospective analysis, we detect BoDV-1 RNA 1 day after hospital admission when the cell count in the cerebrospinal fluid is still normal. We develop a new ELISA using recombinant BoDV-1 nucleoprotein, phosphoprotein, and accessory protein X to detect seroconversion on day 12. Antibody responses are also shown in seven previously confirmed cases. The individual BoDV-1 antibody profiles show variability, but the usage of three different BoDV-1 antigens results in a more sensitive diagnostic tool. Our findings demonstrate that early detection of BoDV-1 RNA in cerebrospinal fluid and the presence of antibodies against at least two different viral antigens contribute to BoDV-1 diagnosis. Physicians in endemic regions should consider BoDV-1 infection in cases of unclear encephalopathy and initiate appropriate diagnostics at an early stage.

Human Borna disease virus 1 (BoDV-1) encephalitis cases in the north and east of Germany.

In 2021, three encephalitis cases due to the Borna disease virus 1 (BoDV-1) were diagnosed in the north and east of Germany. The patients were from the states of Thuringia, Saxony-Anhalt, and Lower Saxony. All were residents of known endemic areas for animal Borna disease but without prior diagnosed human cases. Except for one recently detected case in the state of Brandenburg, all >30 notified cases had occurred in, or were linked to, the southern state of Bavaria. Of the three detected cases described here, two infections were acute, while one infection was diagnosed retrospectively from archived brain autopsy tissue samples. One of the acute cases survived, but is permanently disabled. The cases were diagnosed by various techniques (serology, molecular assays, and immunohistology) following a validated testing scheme and adhering to a proposed case definition. Two cases were classified as confirmed BoDV-1 encephalitis, while one case was a probable infection with positive serology and typical brain magnetic resonance imaging, but without molecular confirmation. Of the three cases, one full virus genome sequence could be recovered. Our report highlights the need for awareness of a BoDV-1 etiology in cryptic encephalitis cases in all areas with known animal Borna disease endemicity in Europe, including virus-endemic regions in Austria, Liechtenstein, and Switzerland. BoDV-1 should be actively tested for in acute encephalitis cases with residence or rural exposure history in known Borna disease-endemic areas.

Borna disease virus in Raccoons (Procyon lotor) in Japan.

We have examined the seroprevalence of BDV in wild Raccoons (Procyon lotor) in Hokkaido, Japan. Serum samples from raccoons were examined using ELISA and Western blot assays to detect the presence of serum antibodies that react specifically to BDV antigens. Among 549 investigated individuals, eleven (2.0%) showed a positive reaction to BDV antigens. Brain tissue samples from five individuals were subjected to RT-PCR, which detected BDV sequences in three of them. Sequence analysis revealed a high degree of genetic conservation between BDV sequences derived from raccoons and previously published sequences derived from other animal species.

Development of a real-time RT-PCR assay for improved detection of Borna disease virus.

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.

Borna disease virus (BDV) infection in cats. A concise review based on current knowledge.

Persistent viral infections of the central nervous system have been the subject of intense interest for decades. One of these viral agents has been identified as Borna disease virus (BDV) of the family Bornaviridae. There have been various reports that link BDV to staggering disease in cats, with symptoms that include ataxia and behavioural disorders, and the disease is often referred to as feline Borna disease. Serological and molecular detection of BDV has been reported at a higher prevalence in cats with neurological disorders in comparison to healthy cats. The transmission route(s) of BDV remain largely unknown, and the hypothesis that BDV is a zoonotic agent is yet to be proven. This review summarises the current knowledge on BDV infection in cats and discusses epidemiological aspects of infection.

Borna disease virus and neuropsychiatric disease--a reappraisal.

Despite progress in understanding the molecular biology and pathobiology of Borna disease virus, its epidemiology and role in human disease remain controversial. The challenges encountered in this field are a paradigm for the investigation of diseases potentially linked to complex host-microorganism interactions.

Antibodies to borna disease virus in subacute sclerosing panencephalitis.

Mechanisms causing persistence and reactivation of measles virus in subacute sclerosing panencephalitis (SSPE) are unknown. Borna disease virus (BDV) frequently causes latent or persistent infection in the nervous system. We investigated a possible association of these viruses in SSPE. Although BDV seropositivity was similar in SSPE and control groups, SSPE patients with high antibodies to BDV had earlier and more rapid disease. The findings suggest that BDV might be involved in the course, but not in the etiopathogenesis, of SSPE.

Discrepancy in the diagnosis of avian Borna disease virus infection of Psittaciformes by protein analysis of feather calami and enzyme-linked immunosorbent assay of plasma antibodies.

The present study compares diagnosis of avian Borna disease virus (ABV) infection of psittacine birds by Western blot of bornaviral proteins in dried feather stems with the detection of anti-bornaviral protein antibodies to bornaviral proteins in plasma by enzyme-linked immunosorbent assay (ELISA). The detection of ABV proteins P40 and P24 in feather calami by Western blotting was possible even after storage of the dried feathers for several years at ambient temperature. Serological identification of anti-bornaviral antibodies may fail (e.g., in young birds, hatched from infected parents), whereas bornaviral P40 and P24 proteins were detected in feather stems. This failure can last at least 10 months after the birds are hatched. In some older birds (>5 years), ABV protein was only detectable in the brain, but not in some peripheral tissues, suggesting that the immune system had succeeded in removing the infecting ABV from tissues outside the brain. These results show that a combination of feather stem analysis for the presence of bornaviral proteins by Western blot combined with serological detection of anti-bornaviral antibodies by ELISA is the most reliable procedure for the detection of a bornaviral infection.

Borna disease virus infection in cats: ABCD guidelines on prevention and management.

OVERVIEW: Borna disease virus (BDV) has a broad host range, affecting primarily horses and sheep, but also cattle, ostriches, cats and dogs. In cats, BDV may cause a non-suppurative meningoencephalomyelitis ('staggering disease'). INFECTION: The mode of transmission is not completely elucidated. Direct and indirect virus transmission is postulated, but BDV is not readily transmitted between cats. Vectors such as ticks may play a role and shrews have been identified as a potential reservoir host. Access to forested areas has been reported to be an important risk factor for staggering disease. DISEASE SIGNS: It is postulated that BDV may infect nerve endings in the oropharynx and spread via olfactory nerve cells to the central nervous system. A strong T-cell response may contribute to the development of clinical disease. Affected cats develop gait disturbances, ataxia, pain in the lower back and behavioural changes. DIAGNOSIS: For diagnostic purposes, detection of viral RNA by reverse transcription PCR in samples collected from cats with clinical signs of Borna disease can be considered diagnostic. Serology is of little value; cats without signs of Borna disease may be seropositive and yet not every cat with BDV infection has detectable levels of antibodies. HUMAN INFECTION: A hypothesis that BDV infection may be involved in the development of selected neurological disorders in man could not be confirmed. A research group within the German Robert Koch Institute studied the potential health threat of BDV to humans and concluded that BDV was not involved in the aetiology of human psychiatric diseases.

High-avidity human serum antibodies recognizing linear epitopes of Borna disease virus proteins.

BACKGROUND: The recent observation that Borna disease virus (BDV)-reactive antibodies from psychiatric patients exhibit only low avidity for BDV antigen called into question their diagnostic value and raised the possibility that antigenically related microorganisms or self antigens caused the production of these antibodies. We further characterized the specificity of these antibodies. METHODS: We established a peptide array-based screening test that allows the identification of antibodies directed against linear epitopes of the two major BDV proteins, the nucleoprotein (N) and the phosphoprotein (P). RESULTS: Initial tests employing sera of BDV-infected mice and rats or horses with Borna disease revealed a high specificity and sensitivity of this test. All sera recognized epitopes of N, P, or both. Sera of noninfected rats, mice, and horses showed no signals on either peptide array. Several human sera that recognized BDV antigen by indirect immunofluorescence contained antibodies that recognized various linear epitopes of one or even both BDV proteins. Remarkably, antibodies purified from such human serum by matrix-immobilized peptides showed high-avidity binding to BDV antigens when assayed by IFA or Western blotting. CONCLUSIONS: These data suggest that reactive antibodies found in psychiatric patients indeed indicate infection with BDV or a BDV-like agent. However, the poor affinity maturation of BDV-specific human antibodies remains unexplained.

Borna disease virus infection in psychiatric patients: are we on the right track?

Animals infected with Borna disease virus (BDV) typically present with neurological dysfunction including behavioural abnormalities. Seroepidemiological surveys suggested that BDV infection can occur in human beings and is associated with mental disorders. Partly contradictory results from studies employing RT-PCR and serological screening led to debate over whether BDV can infect people at all. Critical evaluation of available data led to doubts about the diagnostic value of RT-PCR-based test results. A more consistent picture has emerged from serological studies because seropositive cases were found more frequently among psychiatric patients than among normal controls, supporting the notion that BDV might indeed be responsible for some psychiatric disorders. This view is now challenged by the observation that human BDV-reactive antibodies are of low avidity and might therefore represent cross-reacting antibodies. It remains to be shown whether these antibodies are indeed induced by BDV or by related antigens of unknown identity.

Borna disease virus and infection in humans.

Borna disease virus (BDV) is a nonsegmented, negative-, single-stranded, highly neurotropic RNA virus with noncytolytic replication in the central nervous system. This virus causes neurological and behavioral disturbances primarily in horses and sheep, in addition to a variety of other vertebrate animal species and in laboratory animal models. BDV is now gaining much of the research attention, because the disturbances seen in animals resemble those of neuropsychiatric disorders in humans. These observations raise the possibility that BDV infection may be associated with certain human disorders. Serological and molecular studies on many samples from human patients with a variety of psychiatric disorders have been performed. Some reported the presence and elevated levels of serum antibodies to BDV. Others reported the presence of BDV-RNAs or BDV-antigens in the peripheral blood samples as well as in autopsied brains. Taken together these data support the possibility of human infection with BDV. On the contrary, others reported the complete absence of such BDV-markers from their samples, supporting the absence of a link between BDV infection and psychiatric disorders as well as excluding it as a human pathogen. Thus, BDV infection in humans is highly controversial. Further investigations are required to answer the question whether BDV is a human pathogen and moreover, to elucidate the possible role, if any, of BDV in the pathogeneses of these disorders.

Varied prevalence of Borna disease virus infection in Arabic, thoroughbred and their cross-bred horses in Iran.

Borna disease virus (BDV) naturally infects horses and sheep and induces progressive poliomeningoencephalomyelitis. Here, BDV recombinant proteins of the first open reading frame (ORF-I; coding for p40 nucleoprotein) and the second ORF-II (coding for p24 polymerase cofactor) were immunoblotted with plasma derived from 72 healthy (28 Arabic, 17 thoroughbred and 27 cross-bred) race horses at Tehran in Iran to detect anti-BDV antibodies. In addition, their peripheral blood mononuclear cells (PBMCs) were also examined for BDV RNA by a nested reverse transcriptase-polymerase chain reaction (RT-PCR) at ORF-II. The prevalence of BDV antibodies and/or RNA was 41.2% in Arabic, 23.5% in thoroughbred, and 33.3% in cross-bred horses, but only 17.9, 5.9, and 11.1% of them, respectively, showed positive signals for both BDV antibodies and RNA. Especially, cross-bred horses showed a higher prevalence for BDV RNA, which was detected only in females. In addition, significantly higher prevalence for BDV RNA was observed in Arabic males and thoroughbred females. The BDV prevalence did not increase with aging of the horse. Sequencing at the region of BDV derived from Iranian horses revealed a slight difference from those of Japanese horse- and European horse-derived BDVs even in the amino acid residues, although those in the three groups of Iranian horses were quite similar. Thus, the varied prevalence of BDV was observed with the horse strain or sex in Iranian horses, although BDV sequences were very similar among all three groups in Iran compared with those derived from other countries.

Borna disease virus--does it infect humans and cause psychiatric disorders?

Antibodies recognizing Borna disease virus (BDV) antigens were first demonstrated in the blood of psychiatric patients approximately 15 years ago. Since that time, a highly controversial debate arose whether BDV infects humans and whether it causes psychiatric disorders. In this review, we critically discuss the results of numerous studies that assessed this possibility by using virological and serological methods. We conclude that there is presently no strong experimental evidence supporting the notion that BDV is a human pathogen. The possibility remains, however, that an antigenically related agent is associated with human psychiatric disorders.

Sensitivity and reproducibility of RT-PCR to detect Borna disease virus (BDV) RNA in blood: implications for BDV epidemiology.

Borna disease virus (BDV) infection of domestic animals and humans appears to have a worldwide distribution. There is evidence suggesting an association of BDV with certain psychiatric disorders. However, more comprehensive epidemiological studies are required to establish rigorously a link between BDV and human mental disorders, and to evaluate the role of carrier animals as potential source of BDV for human infection. The use of RT-PCR to detect BDV RNA in peripheral blood mononuclear cells (PBMCs) of infected individuals is a powerful tool to address these questions. The comparison of discrepant results reported by different investigators using this approach is hampered by the lack of controls to assess the sensitivity and reproducibility of the assays. Procedures are now described that allow the establishment of standardized controls to evaluate the performance of the RT-PCR assays. This RT-PCR assay detected reproducibly 100 copies of BDV p40 RNA in 5 microg of RNA. The data illustrate that the number of PBMCs used for RNA preparation, rather than the amount of RNA, has a critical influence on the outcome of the RT-PCR assay. Evidence is provided that levels of BDV in blood do not necessarily reflect viral load in brain.

Detection of Borna disease virus genome in normal human brain tissue.

Borna disease virus (BDV), a neurotropic virus naturally infecting horses and sheep, has been suggested to be associated with human psychiatric disorders. Thus far no extensive studies have been done, providing the evidence of BDV genome in normal human brain tissue. We therefore examined four brain regions of 30 normal autopsy brains for BDV p24 genome. By highly sensitive nested reverse transcriptase (RT)-mediated PCR analysis, we found positive PCR products in two brains: one in frontal and temporal cortices and hippocampus and another in frontal cortex and olfactory bulb. Our results suggest that BDV can infect human brain tissue latently, without causing an apparent neuropsychiatric disorder.