牛传染性鼻气管炎病毒检测方法研究进展

牛传染性鼻气管炎病毒是牛易感的病毒性传染病,以呼吸道症状为主,是一种急性、热性、接触性传染病。近年来,牛传染性鼻气管炎病毒的发病呈上升趋势,对养牛产业危害较大,加强牛传染性鼻气管炎病毒的实验室检测对预防和控制本病起到关键作用,本文主要综述牛传染性鼻气管炎病毒的检测方法研究进展,为相关疾病的诊断及防控提供参考。     

牛传染性鼻气管炎病毒内蒙古分离株gG基因的PCR扩增

参考牛传染性鼻气管炎病毒全基因序列(GenBank)设计1对特异性引物,以牛传染性鼻气管炎病毒内蒙古分离株提取的总DNA为模板,运用PCR方法成功地扩增出牛传染性鼻气管炎病毒内蒙古分离株gG基因,并用琼脂糖凝胶电泳检测扩增产物。     

宁夏地区奶牛病毒性腹泻及牛传染性鼻气管炎的流行病学调查

为了解宁夏地区奶牛的病毒性腹泻和传染性鼻气管炎的流行情况,本研究对宁夏不同地区的奶牛采取了135份耳组织和376份血清,分别对牛病毒性腹泻病毒抗原及牛传染性鼻气管炎病毒抗体进行了检测。结果显示,牛病毒性腹泻病毒抗原阳性率最高为0.5%,平均阳性率为0.01%;牛传染性鼻气管炎病毒抗体阳性率最高为100%,平均阳性率为85.1%。证实了宁夏地区已经存在着较为严重的牛病毒性腹泻病毒及牛传染性鼻气管炎病毒的感染。牛病毒性腹泻以及牛传染性鼻气管炎的流行病学调查为我区制定出科学有效的综合防控措施及促进地区奶业健康可持续发展提供依据。     

牛传染性鼻气管炎病毒抗体胶体金检测试纸条的制备

采用SF 9昆虫细胞-杆状病毒系统重组表达牛传染性鼻气管炎病毒gD蛋白抗原,经纯化鉴定后将gD蛋白用胶体金标记作为示踪抗原,未标记的gD抗原作为捕获抗原,并以羊抗牛IgG抗体作为质控抗体,建立了牛传染性鼻气管炎病毒双抗原夹心法胶体金检测试纸条。试验结果表明,该试纸条检测灵敏度高,特异性良好,与牛口蹄疫病毒、牛病毒性腹泻病毒等阳性血清无交叉反应。使用建立的胶体金试纸条和IDEXX牛传染性鼻气管炎病毒抗体ELISA检测试剂盒同时检测112份牛血清,阳性符合率为93.5%,阴性符合率为96.0%,总符合率为94.6%。说明该试纸条可以应用于临床牛传染性鼻气管炎病毒抗体的诊断和检测。     

牛传染性鼻气管炎诊断方法的研究进展

牛传染性鼻气管炎是由牛传染性鼻气管炎病毒引起的一种牛的热性、急性、接触性传染病。该病是世界动物卫生组织(OIE)规定的必须上报的疾病之一,在我国也被列为二类疫病。牛传染性鼻气管炎可降低牛的肥育率、繁殖率和产奶量,给养牛业造成了重大经济损失。从病毒的分离鉴定、血清学以及分子生物学等方面对该病的诊断方法进行综述,以期为牛传染性鼻气管炎的检测和防控提供参考。     

牛传染性鼻气管炎诊断方法研究进展

牛传染性鼻气管炎是由牛传染性鼻气管炎病毒引起的一种牛的热性、急性、接触性传染病。该病是世界动物卫生组织(OIE)规定的必须上报的疾病之一,在我国也被列为二类疫病。牛传染性鼻气管炎可降低牛的肥育率、繁殖率和产奶量,给养牛业造成了重大经济损失。从病毒的分离鉴定、血清学以及分子生物学等方面对该病的诊断方法进行综述,以期为牛传染性鼻气管炎的检测和防控提供参考。     

我国部分地区牛传染性鼻气管炎血清学调查与分析

为了解我国牛传染性鼻气管炎的感染情况及流行趋势,对2014年-2016年采集于11个牧区半牧区省份的9 668份牛血清样品进行了牛传染性鼻气管炎病毒抗体检测。结果显示,牛传染性鼻气管炎病毒抗体的个体平均阳性率为52.77%,群体阳性率连续3年均在80%以上,说明感染现象较为普遍。乳用牛的感染情况较肉用牛和乳肉兼用牛严重。同时,发现生产实际中牛传染性鼻气管炎与牛病毒性腹泻混合感染情况普遍。此次摸底调查掌握了我国牛传染性鼻气管炎感染情况,为制定科学防控政策提供了参考依据。     

牛传染性鼻气管炎的诊断与防制

牛传染性鼻气管炎又称坏死性鼻炎、红鼻病,是由牛传染性鼻气管炎病毒引起的牛的一种接触性传染病,在临床上以呼吸道、支气管黏膜发炎、呼吸困难、流鼻液等为特征,还可出现生殖道感染、结膜炎、脑膜炎、流产、乳房炎等临床表现.目前,本病世界范围内流行,给全球的的养牛业带来极大影响.我国规定,进口牛及其肉制品必须检测牛传染性鼻气管炎病毒.     

新疆部分地区牛传染性鼻气管炎病毒感染的检测与gD基因序列分析

为了调查新疆部分地区牛传染性鼻气管炎病毒的感染情况,试验从新疆石河子、奎屯、库尔勒、沙湾、阿克苏地区的五个规模化奶牛场采集1月龄以内犊牛鼻液126份,其中发病犊牛86份,相同日龄健康犊牛40份,采用双抗体夹心ELISA方法检测牛传染性鼻气管炎病毒抗原,PCR方法检测牛传染性鼻气管炎病毒g D基因,并挑选不同地区的9株病毒进行序列分析比较其同源性。结果表明:ELISA方法检测的感染率为19.84%,PCR方法检测的感染率为47.62%,g D基因序列同源性为77.80%~99.80%。说明新疆部分地区牛传染性鼻气管炎病毒感染较为普遍,且多数毒株之间基因变异较大。     

牛传染性鼻气管炎病毒SYBR GreenⅠ荧光定量PCR检测方法的建立

为检测牛传染性鼻气管炎病毒,通过设计针对gB基因的特异性引物扩增gB基因,构建阳性重组质粒。经过优化建立了牛传染性鼻气管炎病毒的SYBR GreenⅠ荧光定量PCR检测方法。结果显示,建立的实时荧光定量PCR检测方法在阳性标准质粒拷贝数在5.474×10⁴～5.474×10⁹拷贝/μL时线性关系良好,线性相关系数R²=0.999 3。该方法最低检测限为5.747×10²拷贝/μL;可以特异性检测牛传染性鼻气管炎病毒(IBRV),与牛病毒性腹泻病毒(BVDV)、牛副流感病毒(BPIV)、牛呼吸道合胞体病毒(BRSV)不发生交叉反应。病毒滴度与拷贝数的线性关系良好,线性相关系数R²=0.985 4。该方法为牛传染性鼻气管炎病毒的快速检测提供了技术支持。     

牛传染性鼻气管炎诊断方法研究进展

牛传染性鼻气管炎(IBR)是由牛传染性鼻气管炎病毒(IBRV),即牛疱疹病毒1型(BoHV-1)所引起的以上呼吸道炎症为主的一种牛的急性、热性、接触性传染病,呈世界性流行。IBR的早期准确诊断,对该病的防控具有不可忽视的作用。目前,IBR的诊断方法主要包括病原学诊断和血清学诊断方法。病原学诊断具有特异和敏感及准确等特点,而血清学诊断具有敏感、快速、方便和价廉等特点。为了实施IBR的净化和根除计划,部分国家和地区已逐渐采用IBR基因缺失疫苗,配套使用鉴别诊断方法来鉴别IBR疫苗免疫和自然感染。论文就牛传染性鼻气管炎常用诊断方法的研究进展进行综述,以期为IBR的诊断和防控提供参考。     

In vitro interferon production by bovine tissues: induction with infectious bovine rhinotracheitis virus.

The interferon-inducing ability of infectious bovine rhinotracheitis (IBR) virus was determined in tissue cultures of bovine origin inoculated with untreated and ultraviolet (UV) irradiated IBR viruses. Interferon was assayed by the plaque-reduction method in bovine fetal kidney (BFK) cell cultures, using vesicular stomatitis virus as challenge virus. Highest interferon concentrations were produced by cultures of bovine fetal (BF) spleen cells and aveolar macrophage cultures derived from adult cattle. Moderate interferon concentrations were produced by peripheral blood leukocyte (PBL) suspension cultures from adult cattle with serum-neutralizing antibodies against IBR virus. Cultures of PBL from 1 cow without detectable serum-neutralizing antibodies against IBR virus did not produce detectable interferon in response to IBR virus. Cultures of PBL from cattle with or without detectable serum-neutralizing antibodies against IBR virus produced interferon when stimulated with phytohemagglutinin (PHA). Low levles of viral inhibitors were detected infrequently in monolayer cultures of BFK and BF nasal mucosa inoculated with UV-irradiated IBR virus and in BF tracheal organ cultures inoculated with untreated IBR virus. Interferon was not detected in fluids collected from IBR virus-exposed monolayer cultures of primary and secondary BF lung, secondary BF tracheal mucosa, secondary BF liver, secondary BF adrenal, and PBL in the 4th and 7th passages. The antiviral inhibitors from BF spleen, bovine alveolar macrophage, and PBL cultures induced with IBR virus, as well as inhibitors from PBL cultures induced with PHA, had the usual properties of interferon.     

Enzyme linked immunosorbent assay used to monitor serum antibodies to bovine respiratory disease viruses

An enzyme linked immunosorbent assay (ELISA) was applied to the detection of serum antibodies against infectious bovine rhinotracheitis (IBR), parainfluenza-3 (PI3), adenovirus type 3 (adeno 3) and bovine respiratory syncytial (BRS) viruses. Paired serum samples from calves vaccinated with live attenuated virus vaccines were tested. The ELISA compared favorably with the virus neutralization test for detecting serologic responses to IBR, BRS, and adeno 3 viruses or with the hemagglutination inhibition test for PI3 virus. The simplicity, sensitivity and rapidity of the ELISA test makes it a useful tool for immunological studies with respiratory viruses.     

Periocular dermoid cyst in a calf

Serum samples (n = 1,146) representing 100 species of exotic ruminants now captive in United States zoos were assayed for neutralizing antibody to infectious bovine rhinotracheitis (IBR) virus (bovine herpesvirus 1). Thirty-four animals (3%) of 11 species had antibody to IBR virus. Because of the low prevalence of IBR antibody found, it was concluded that vaccination against IBR virus probably is not necessary for captive wild ruminants in United States zoos.     

A Comparison of the Viruses of Infectious Bovine Rhinotracheitis (IBR), Infectious Pustular Vulvovaginitis (IPV), and Rinderpest: Part I. Studies of Antigenic Relationships

The viruses of infectious bovine rhinotracheitis (IBR), infectious pustular vulvovaginitis (IPV), and rinderpest were compared by specific methods. The results further confirmed that IBR and IPV are caused by agents with common antigens. No antigenic relationship was found between these viruses and rinderpest virus, which confirms earlier work.     

Investigation of causative agents of bovine respiratory tract disease in a beef cow-calf herd with an early weaning program.

Serum samples were collected from early weaned fall calves shortly after the onset of respiratory tract disease. Antibody titers to infectious bovine rhinotracheitis (IBR) virus, parainfluenza type 3 (PI-3) virus, bovine viral diarrhea (BVD) virus, bovine adenovirus type 3 (BAV-3), and bovine respiratory syncytial virus (BRSV) were determined on paired (acute and convalescent) serums. Seroconversion rate (a fourfold or greater rise in antibody titer) for IBR virus was 4.3%, PI-3 virus--16.3%, BVD virus--9.6%, and BAV-3--2.2%. Seroconversion for BRSV was 45.4%. An increased rate of seroconversion for IBR, PI-3, and BVD viruses and BAV-3 was observed in the presence of BRSV seroconversion. These results suggest that BRSV may facilitate infection by other viruses. Results of virus isolation procedures from these calves were negative.     

Experimental transmission of bovine viral diseases by insemination with contaminated semen or during embryo transfer

Three experimental approaches were used to study transmission of blue tongue (BT), infectious bovine rhinotracheitis (IBR) and bovine virus diarrhoea (BVD) viruses. These were insemination with contaminated semen, experimental infection of embryo donor cows, or transfer of embryos experimentally exposed to virus in vitro to normal recipients. Parameters assessed included number and quality of embryos produced, virus detection (isolation and electron microscopy), serology and histopathology. All superovulated sesceptible cows inseminated with semen containing blue tongue virus (BTV) (n = 2) or infectious bovine rhinotracheitis virus (IBRV) (n = 2) became infected. One cow inseminated with semen containing BTV produced seven virus-free seven-day-old embryos; the second cow failed to produce any embryos. One of two cows inseminated with semen containing IBRV produced two underdeveloped, virus-free embryos while no embryos were produced by the second cow. One of two cows inseminated with semen containing bovine viral diarrhoea virus (BVDV) became infected. Two poorly developed, virus-free seven-day-old embryos were recovered from one of these cows. Superovulated susceptible cows inoculated either intramuscularly with BTV (n = 3) or intranasally with IBR virus (n = 2) became infected. Virus was isolated from some tissues of two BTV-infected cows, neither of which produced embryos. A third BTV-infected cow produced two virus-free embryos collected at necropsy five days after inoculation. One of two cows experimentally infected with IBR virus, produced three embryos but virus was not detected either by electron microscopy (1 embryo) or in cell culture by cytopathic alterations (1 embryo).(ABSTRACT TRUNCATED AT 400 WORDS)     

Effects in calves of mixed infections with bovine viral diarrhea virus and several other bovine viruses

The objective of this study was to verify whether a mixed infection in calves with bovine viral diarrhea virus (BVDV) and other bovine viruses, such as bovid herpesvirus-4 (BHV-4), parainfluenza-3 (PI-3) and infectious bovine rhinotracheitis (IBR) virus, would influence the pathogenesis of the BVDV infection sufficiently to result in the typical form of mucosal disease being produced.

Accordingly, two experiments were undertaken. In one experiment calves were first infected with BVDV and subsequently with BHV-4 and IBR virus, respectively. The second experiment consisted in a simultaneous infection of calves with BVDV and PI-3 virus or BVDV and IBR virus.

From the first experiment it seems that BVDV infection can be reactivated in calves by BHV-4 and IBR virus. Evidence of this is that BVDV, at least the cytopathic (CP) strain, was recovered from calves following superinfection. Moreover, following such superinfection the calves showed signs which could most likely be ascribed to the pathogenetic activity of BVDV. Superinfection, especially by IBR virus, created a more severe clinical response in calves that were initially infected with CP BVDV, than in those previously given the non-cytopathic (NCP) biotype of the virus. Simultaneous infection with PI-3 virus did not seem to modify to any significant extent the pathogenesis of the experimentally induced BVDV infection whereas a severe clinical response was observed in calves when simultaneous infection was made with BVDV and IBR virus.     

Detection of bovine rhinotracheitis virus antibody by neutralizing test and ELISA in experimentally infected rabbits.

The antibody response of rabbit to infectious bovine rhinotracheitis (IBR) virus was examined. Two rabbits were inoculated with IBR virus strain Los Angeles into the trachea and intravenously, and intravenously two times, respectively. The patterns of antibody titers in the rabbits measured by neutralization test and ELISA were similar to those in the case of bovine. The antibody was detected after the inoculation, and much more antibody was detected after the second inoculation. The fact suggests that rabbits are a very useful laboratory animal for IBR virus infection studies.     

Evaluation of a quadrivalent inactivated vaccine for the protection of cattle against diseases due to common viral infections

Efficacy of an inactivated quadrivalent vaccine containing infectious bovine rhinotracheitis (IBR) virus, parainfluenza type 3 (PI3) virus, bovine virus diarrhoea virus (BVDV) and bovine respiratory syncytial virus (BRSV) was assessed in naive bovine calves to evaluate short-term (4-18 weeks) and long-term (24-38 weeks) protection following the basic intramuscular vaccination regime of 2 inoculations a month apart. Vaccination was staggered between the long-term and the short-term groups by about 5 months so that both groups, along with a matched group of 6 unvaccinated (control) calves, could be challenged at the same time. Sequential challenges at intervals of 3-8 weeks were done in the order: IBR virus (intranasally, IN), PI3 virus (IN and intratracheally, IT), pestiviruses (IN) and BRSV (IN and IT). The IBR virus challenge produced febrile rhinotracheitis (FRT) in control calves but both the severity and the duration of FRT was significantly reduced in both vaccinated groups. The amount and the duration of IBR virus shed by the vaccinated groups was significantly reduced compared to the control group. Although PI3 virus, pooled pestivirus and BRSV challenges did not result in a noteworthy disease, challenge virus shedding (amount and duration) from the upper (all 3 viruses) and the lower (BRSV) respiratory tracts was significantly reduced in vaccinated groups. After pestivirus challenge, sera and leukocytes from all control calves were infectious for 6-9 days whereas virus was recovered only from leukocytes in vaccinated calves and only for 1.6-2.7 days. Thus a standard course of the quadrivalent vaccine afforded a significant protection against IBR virus, PI3 virus, BVDV and BRSV for at least 6 months.