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1.
A blocking enzyme-linked immunosorbent assay (ELISA) has been adapted to detect specific antibodies in bovine sera to respiratory syncytial virus using a horseradish peroxidase-labeled monoclonal antibody to the fusion protein of the virus. This assay plus an indirect blocking ELISA and indirect ELISA were used to detect antibodies to the bovine respiratory syncytial virus (BRSV) in 159 field-origin bovine sera. Results of these assays were compared with serum antibody titers measured by the serum neutralization (SN) test. Over a 56-day period, the mean neutralization titers and the mean delta absorbance values for the blocking ELISA, on the same sera, showed similar declines. However, the calculated correlation coefficients between mean SN titer and mean absorbance value for the blocking ELISA of the individual sera ranged from -0.2 to -0.5 depending on the source of sera. Similar values were obtained whether using crude or purified viral antigen in the assays. Corresponding calculated correlation coefficients were generally higher for the indirect blocking ELISA or indirect ELISA than for the blocking ELISA. The blocking ELISA was between 70 and 64% as sensitive as the serum neutralization test with a specificity of 100 or 90% using the crude and purified viral antigen, respectively. The indirect blocking ELISA and indirect ELISA had similar calculated sensitivities and specificities. The blocking ELISA was faster to run than either of the other ELISA's or the neutralization test. Further, nonspecific background absorbance was obviated because the blocking ELISA detects antibodies to 1 specific viral protein, the fusion protein. These studies suggest that the blocking ELISA should be useful as a serological test for BRSV antibodies.  相似文献   

2.
Respiratory syncytial virus (RSV) causes severe lower respiratory tract infection in children and calves. Antibodies to ovine RSV (ORSV) are common in sheep, but the clinical disease is not well defined. There is no report of ORSV infection in Australian sheep although respiratory distress syndrome has been described. This discrepancy may be due to the lack of a suitable diagnostic test. In this report, we have characterised the ORSV G protein in an attempt to study its relatedness to human and bovine RSV (HRSV, BRSV) and for use in the development of a suitable diagnostic assay. Full length and a truncated variant of ORSV G protein were expressed in mammalian cells and the expressed proteins characterised by indirect immunofluorescence and radioimmunoprecipitation assays. Our results indicate that like HRSV, the ORSV G protein is heavily glycosylated. The expressed protein was membrane bound as well as secreted and could be purified from culture supernatants and may be suitable for use in development of a diagnostic assay.  相似文献   

3.
An indirect double antibody sandwich enzyme-linked immunosorbent assay (ELISA) was developed for the detection and titration of serum antibodies to bovine respiratory syncytial virus (BRSV). The ELISA was compared with a complement fixation (CF) test and a test for virus neutralising antibody in serum (virus neutralisation [VN] test). Testing sera collected in dairy herds revealed the closest correlation between the results of the ELISA and the CF test with respect to BRSV antibody titres. The VN test detected BRSV antibodies in a higher percentage of acute phase sera compared to the other two tests in field samples and in early bleedings of experimentally infected calves. However, the VN test was less effective in making a diagnosis of BRSV infections on the basis of a significant titre increase in paired sera. For this purpose the ELISA was found to be the most sensitive test.  相似文献   

4.
Seroprevalence of bovine respiratory syncytial virus (BRSV) infection in both exotic and crossbred cattle were described. A baculovirus expressed recombinant purified nucleocapsid (N) protein was used in indirect and sandwich ELISA for screening of 499 bovine sera samples from all over the state for the presence of BRSV antibodies. The seroprevalence rate of BRSV was found to be 46.09% through indirect ELISA while it would found to be 65.33% by sandwich ELISA. The result also indicated that exotic breeds were more susceptible to BRSV infection compared to crossbred cattle. A comprehensive analysis on susceptibility to BRSV as regards to various factors like age and sex was also summarized.  相似文献   

5.
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6.
Human and bovine respiratory syncytial virus: immunopathologic mechanisms   总被引：2，自引：0，他引：2  
Human respiratory syncytial virus (HRSV) is the major respiratory tract pathogen of infants and young children. Bovine respiratory syncytial virus (BRSV) is recognised as an important cause of respiratory tract disease in calves. Both of these viruses and their respective diseases share many similarities. Immunopathologic mechanisms have been proposed to be involved in the pathogenesis of respiratory syncytial virus (RSV) infections. This review examines the current understanding of the role of immunopathologic mechanisms in RSV infections. The role of vaccines in inducing hypersensitivity is also examined. Additionally, non-immunopathogenic mechanisms involved in RSV infections are discussed.  相似文献   

7.
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8.
Virus shedding was monitored in nasal secretions of 12 calves experimentally infected with bovine respiratory syncytial virus (BRSV) using an antigen capture enzyme-linked immunosorbent assay (ELISA) detecting the nucleoprotein (NP) antigen of BRSV, by a polymerase chain reaction (PCR) amplifying the fusion protein of BRSV, and by a microisolation assay combined with immunoperoxidase staining for the F protein of BRSV. Under the conditions of this study, similar limits of detection and quantitative results were obtained from all three assays. BRSV was detected in nasal secretions of all calves for a minimum of 4 d. Virus shedding began on Day 2 after infection, peaked on Days 3-5, and was cleared in most calves by Day 8. The PCR, and to a lesser extent the ELISA, may detect virus shedding for a longer period after infection than virus isolation, possibly due to neutralization of the virus by rising mucosal antibody. Simulated environmental conditions likely to be experienced during transport of clinical field specimens markedly reduced the sensitivity of virus isolation but had a minimal effect on the results of the NP ELISA. Actual field transport conditions (overnight on ice) had minimal apparent effect on the results of the PCR assay. The less stringent specimen handling requirements, combined with low limits of detection, of both the nucleoprotein ELISA and PCR, indicate either of these assays are more suitable for diagnostic applications than virus isolation.  相似文献   

9.
In a group of 60 Belgian White Blue calves less than 8 months old still housed in barns, a bovine respiratory syncytial virus (BRSV) outbreak was revealed on the basis of a direct diagnosis (immunofluorescence and virus isolation) performed on the lungs of dead animals, and the kinetics of BRSV neutralizing antibodies. Clinical signs, macroscopical and microscopical pulmonary lesions were also compatible with a BRSV infection. This outbreak is peculiar because the 35 oldest calves (204 +/- 29 days old) had been vaccinated 3-4 months before with an inactivated BRSV vaccine and 30% of these animals had died of respiratory distress. While they experienced a mild respiratory symptomatology, no death was recorded among the 25 youngest calves (69 +/- 29 days old) which had been left unvaccinated. Another peculiarity was found at the histological level where a massive infiltration of eosinophils was demonstrated in the pulmonary tissues of the dead animals. Together these data parallel the dramatic story described 30 years ago in children previously vaccinated with a formalin-inactivated human RSV (HRSV) vaccine upon a natural HRSV challenge. This illustrates that an immunopathological phenomenon also takes place after BRSV vaccination in cattle.  相似文献   

10.
OBJECTIVE: To develop a model of bovine respiratory syncytial virus (BRSV) infection that induces severe disease similar to that seen in some cattle with naturally acquired BRSV infection. ANIMALS: 25 male Holstein calves, 8 to 16 weeks old. PROCEDURE: 17 calves were given a low-passage field isolate of BRSV by aerosolization; 8 control calves were given supernatant from noninfected cell culture. Disease was characterized by evaluating clinical signs, virus isolation and pulmonary function tests, and results of blood gas analysis, gross and histologic postmortem examination, and microbiologic testing. RESULTS: Cumulative incidence of cough, harsh lung sounds, adventitious sounds, and dyspnea and increases in rectal temperature and respiratory rate were significantly greater in infected calves. Three infected calves developed extreme respiratory distress and were euthanatized 7 days after inoculation. Virus was isolated from nasal swab specimens from all infected calves but not from mock infected calves. On day 7 after inoculation, mean PaO2 and PaCO2 were significantly lower, and pulmonary resistance was significantly higher, in infected calves. During necropsy, infected calves had varying degrees of necrotizing and proliferative bronchiolitis and alveolitis with syncytial formation. The 3 calves euthanatized on day 7 had emphysematous bullae in the caudal lung lobes; 1 had unilateral pneumothorax. CONCLUSION AND CLINICAL RELEVANCE: Severe disease similar to that seen in some cattle with naturally acquired BRSV infection can be induced in calves with a single aerosol exposure of a low-passage clinical isolate of BRSV. Our model will be useful for studying the pathogenesis of BRSV infection and for evaluating vaccines and therapeutics.  相似文献   

11.
Development of fluorescence in bovine embryonic lung cells infected with bovine respiratory syncytial virus (BRSV) was studied by the fluorescent antibody (FA) test. Similar patterns of fluorescence were seen with the direct FA test, in which the immunoglobulin G fraction of antiserum to BRSV was conjugated with fluorescein isothiocyanate and used; and the indirect test, in which antiserum to the Long strain of respiratory syncytial virus and fluorescein isothiocyanate-conjugated anti-rabbit immunoglobulin G were used. In different trials, fluorescence was first detected between 16 and 18 hours after inoculation with BRSV. Fluorescence always was confined to the cytoplasm. Before 24 hours, fluorescence consisted of fine fibrils, usually parallel to the long axis of the cell, and cytoplasmic granules. After 24 hours, coincident with rounding of the cells, fluorescence slowly moved to the periphery of the cytoplasm. Under the growth conditions used, syncytia did not develop. By the FA test and as determined by the release of BRSV into the supernatant fluid, the minimal time for a single cycle of infection was between 24 and 26 hours.  相似文献   

12.
A cross-sectional study was carried out, from November 2007 to March 2008, to estimate the prevalence of and to determine risk factors associated with bovine syncytial respiratory virus (BRSV) and parainfluenza 3 virus (PIV3) in dual-purpose herds in Colima, México. One hundred and seventy-six sera from 33 herds for PIV3 and 232 sera from 44 herds for BRSV were used. Sera were analyzed by indirect ELISA for the detection of antibodies against BRSV and PIV3 in cattle herds to determine the seroprevalence of respiratory diseases. The apparent and true prevalences for PIV3 were 60.8% and 54.4% and for BRSV 52.2% and 50.8%, respectively. The percentage of herds showing at least one positive animal was 78.7% for PIV3, and 93.2% for BRSV. Age (≤12, 13-48, and >48?months old) and respiratory signs (no, yes) showed significant association (P?相似文献   

13.
Bovine respiratory syncytial virus (BRSV) is a respiratory pathogen of cattle that causes severe disease in calves alone and as one of several viruses and bacteria that cause bovine respiratory disease complex. Like human RSV this virus modulates the immune response to avoid stimulation of a vibrant CD8+ T cytotoxic cell response and instead promotes a Th2 response. The Th2 skew sometimes results in the production of IgE antibodies and depresses production of the Th1 cytokine interferon γ. Innate immune cells have a pivotal role in guiding the adaptive response to BRSV, with selective secretion of cytokines by pulmonary dendritic cells. Here we review some of the pertinent observations on immune responses to BRSV infection and vaccination and illustrate how experimental infection models have been used to elucidate the immunopathogenesis of BRSV infection. Recent experiments using intranasal vaccination and/or immune modulation with DNA based adjuvants show promise for effective vaccination by the stimulation of Th1 T cell responses.  相似文献   

14.
Cattle inoculated with bovine respiratory syncytial virus (BRSV) were evaluated for the development of a cell-mediated immune response. Results of the leukocyte migration-inhibition test under agarose and the delayed hypersensitivity test indicated that a cell-mediated immune response was elicited after intranasal inoculation of calves with BRSV. Migration inhibition in the leukocyte migration-inhibition test was detected by postinoculation day (PID) 5 and reached maximum inhibition on PID 21. Inhibition of leukocyte migration was still evident by PID 42 when values were still appreciably greater than preinoculation values. All of the calves inoculated with BRSV developed a delayed hypersensitivity skin response when challenge exposed intradermally with BRSV antigen.  相似文献   

15.
Bovine respiratory syncytial virus (BRSV) is a major cause of respiratory disease in calves resulting in a substantial economic loss for the cattle industry worldwide. In order to determine the presence of BRSV in Uruguay, an immunoenzymatic test was set up, using a recombinant BRSV nucleocapsid (N) protein as the antigen. The N protein was produced in Sf9 insect cells by a recombinant baculovirus expressing the N protein. Serum samples collected from one hundred cattle from four different geographic regions of Uruguay were analyzed. Antibodies against the N protein of BRSV were detected in 95% of the serum samples analyzed. These results show for the first time the presence of BRSV antibodies and suggest a widespread BRSV infection in the cattle population of Uruguay.  相似文献   

16.
Severe respiratory disease associated with bovine respiratory syncytial virus (BRSV) infection has been identified in dairy cattle in New York State. The cases identified occurred in dairy calves and heifers. The disease was characterized in 4 animals by pathologic changes including interstitial pneumonia, necrotizing bronchiolitis with multinucleated syncytial epithelial cells and interstitial emphysema. BRSV antigen was demonstrated in lung samples or was isolated in tissue culture in all 4 cases. A retrospective survey of 6279 bovine diagnostic accessions between 1977 and 1982 revealed 66 cases of interstitial pneumonia, often with concurrent bronchiolitis. In this 5 year period, only 1 case in 1981 had interstitial pneumonia and bronchiolitis with pathologic features consistent with BRSV infection. It is concluded that pathogenic BRSV has entered New York State and that it is contributing to clinical respiratory disease in dairy cattle.  相似文献   

17.
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18.
The role of IgE antibodies against respiratory syncytial virus has attracted attention for both human and bovine disease. To detect such antibodies, we have developed an enzyme-linked immunosorbent assay (ELISA) specific for bovine respiratory syncytial virus (BRSV). Firstly, anti-serum strongly positive for BRSV-specific IgE was produced by immunizing a levamisole-treated calf with BRSV. The presence and specificity of BRSV-specific IgE in this animal was confirmed with the Praunitz-Kustner (PK) technique. Potential interference in an ELISA by other BRSV-specific immunoglobulin isotypes was eliminated by preferential precipitation of serum samples with 27.5% saturated ammonium sulfate. The correlation between the PK and the ELISA assay was greater than 93% and the ELISA was found to be more specific than the PK. Indeed, in a pilot experimental infection study, the serum levels of BRSV-specific IgE were found to correlate with the symptom expression following repetitive live virus aerosolization. This may prove to be a useful rapid test to study both herd immunity and the potential pathogenic influence of IgE.  相似文献   

19.
为了检测牛呼吸道合胞体病毒(BRSV),根据已发表的融合蛋白(F)基因序列,设计了套式RT-PCR引物,初步建立了BRSV的套式RT-PCR检测方法。对138份牛鼻腔棉拭子进行了检测,结果检测到了BRSV阳性样品54份,总的阳性检出率为39.1%。对大部分BRSV阳性样品的F基因扩增产物进行了序列测定与分析。用该套式RT-PCR对牛传染性鼻气管炎病毒、牛副流感病毒3型、牛病毒性腹泻病毒等进行了检测,结果无交叉反应,表明该检测方法具有良好的特异性。本研究首次用套式RT-PCR技术证实了我国部分省的牛群中存在BRSV感染。  相似文献   

20.
A serological survey of respiratory virus infections was carried out from 1998 to 2001 in Lecco province, Italy, as part of a health monitoring programme in a population of alpine chamois, many of which died of pneumonia in autumn and winter 2000 to 2001; 194 carcases of all age classes were found over a short period and in a small area. Eighteen of them, which were examined postmortem, consistently showed signs of severe fibrinous lobar pneumonia or catarrhal bronchopneumonia. Samples of serum from 145 chamois collected from hunted animals and carcases were tested by a virus neutralisation test against bovine respiratory syncytial virus (BRSV), bovine viral diarrhoea virus, bovine herpesvirus type 1 and parainfluenzavirus type 3. Positive results were detected only for BRSV. The area was divided into two subunits on the basis of the distribution of deaths; in the areas where fatalities were observed there was a significant increase of BRSV titres at the beginning of the outbreak. Furthermore, during the 2000 and 2001 hunting seasons antibody titres to BRSV were significantly higher in the areas where mortality occurred. The roe deer living in the same area were not affected by pneumonia and had a low prevalence of titres to BRSV which did not vary during the period of the study.  相似文献   

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