布鲁氏菌pcDNA3．1-omp17．3基因疫苗的研制

采用PCR方法扩增了布鲁氏菌17．3ku外膜蛋白编码基因，并将该基因克隆至真核表达载体pcDNA3．1（＋）中，成功构建了真核表达质粒pcDNA3．1-ompl7．3。pcDNA3．1-omp17．3转染coS-7细胞后，通过Western—blotting检测到了17．3ku蛋白的瞬时表达。将pcDNA3．1-ompl7．3免疫小鼠，三免后经ELISA、流式细胞仪以及ELISPOT技术检测到pcDNA3．1-omp17．3在小鼠体内诱导产生了以Th1型为主的细胞免疫应答。结果表明，构建的基因疫苗可作为潜在的布鲁氏菌新型疫苗，有进一步研究的意义。     

Brucella suis in three dogs: presentation,diagnosis and clinical management

Brucella suis is an emerging, zoonotic disease predominantly affecting dogs and humans that engage in feral pig hunting in Australia and other countries. Although B. suis infection in dogs shares some clinical similarities to the host-adapted species (B. canis), B. suis remains an incompletely understood pathogen in dogs with limited published data on its pathogenesis and clinical features. This case series describes the presentations, diagnosis, and clinical management of B. suis infection in three dogs: (1) a bitch with dystocia, abortion and mastitis; (2) an entire male dog with septic arthritis and presumptive osteomyelitis; and (3) a castrated male dog with lymphadenitis. Unique features of these cases are reported including the first documented detection of B. suis from milk and isolation from lymph nodes of canine patients, as well as the follow-up of pups born to a B. suis-infected bitch. Consistent with previous reports, all three dogs showed a favourable clinical response to combination antibiotic therapy with rifampicin and doxycycline. Individually tailored drug regimens were required based on the clinical presentation and other factors, including owner expectations and compliance with therapy as well as a zoonotic risk assessment (generally considered low, except around time of whelping). The authors include their recommendations for the clinical management of dogs that are at-risk or seropositive for B. suis with or without clinical signs or laboratory-confirmed infection.     

呼和浩特地区奶牛流产胎儿中布鲁菌的分离鉴定

布鲁菌病(Brucellosis)是由布鲁菌(Brucella)引起的一种世界范围内的人兽共患传染病,给畜牧业发展带来重大的经济损失。控制布鲁菌病的传播,减少其对人类健康和畜牧业发展的危害已经成为当前亟需解决的问题。该文采用细菌学检验手段,对呼和浩特地区出现流产、早产、死胎、胎盘滞留等疑似布鲁菌病现象的奶牛进行病料采集,并对其进行细菌学分离与鉴定,结果获取了5株布鲁菌菌株,表明该地区存在布鲁菌病。该结果为今后呼和浩特市采取有效措施控制布鲁菌病的流行、蔓延提供了科学依据。     

牛种布鲁杆菌PCR检测方法的研究

为了弥补血清学和微生物学方法检测和诊断布鲁菌病的种种不足,建立用布鲁菌BCSP31聚合酶链反应（PCR）快速检测布鲁菌病原的方法。根据编码牛种布鲁菌31 000外膜蛋白基因序列设计的一对特异性引物,扩增出预期350bp的基因片段。结果,此方法具有较高的特异性和敏感性,DNA最低检测量为0.42pg。结果表明,布鲁菌病原...     

Use of Brucella abortus vaccine strain RB51 in pregnant cows after calfhood vaccination with strain 19 in Argentina

One hundred and seven pregnant cows, which had been calfhood vaccinated with Brucella abortus strain 19 (S-19) were revaccinated with either S-19 or strain RB51 (S-RB51). All S-19-revaccinated animals seroconverted, while none of the RB51-revaccinated animals seroconverted. Two out of 25 (8%) S-19-revaccinated animals aborted, while none of the 57 RB51-revaccinated group aborted. Four of the S-19-revaccinated animals shed S-19 in the milk for at least 7 days, while only 1 cow shed S-RB51 for at least 3 days (but <7 days) post-parturition. Revaccination of strain 19 calfhood-vaccinated, pregnant cattle with S-RB51 appears to be a safe procedure with no diagnostically negative consequences.     

呼和浩特布鲁氏菌分离株bp26基因的原核表达

目的:克隆布鲁氏菌外膜蛋白bp26基因并构建原核表达系统.方法:用聚合酶链式反应技术扩增得到布鲁氏菌bp26基因片段,与PEASY-E1载体链接,转入BL21(DE3)感受态细胞中,经IPTG诱导后,经SDS-PAGE检测重组蛋白表达,用Western blot鉴定目的蛋白的生物活性.结果:DNA测序结果显示,重组质粒bp26基因的插入位点正确,成功构建了重组质粒PEASY-E1-bp26,经IPTG诱导,表达出大小为27kDa的bp26融合蛋白.结论:获得布鲁氏菌外膜蛋白bp26基因片段,并在大肠杆菌中表达出具有生物活性的bp26融合蛋白.     

布鲁菌免疫分子研究进展

布鲁菌病是布鲁菌引起的人兽共患传染病,包括7种21型,其中感染人的主要有牛、羊、猪布鲁菌3种,其免疫涉及体液免疫和细胞免疫.在布鲁菌的免疫过程中,先天性免疫应答主要通过补体、自然杀伤细胞、巨噬细胞、细胞因子等的参与.获得性免疫应答中,CD4 、CD8 、γβT细胞起重要作用.布鲁菌重组亚单位疫苗是近年研究的热点,而且发现的免疫分子也很多.文章围绕布鲁菌免疫机理、免疫相关分子进行探讨,为筛选疫苗候选分子奠定基础.     

布鲁氏菌实时定量荧光PCR检测体系的建立及应用

本根据GenBank上公布的布鲁氏菌BCSP31基因保守区域序列设计合成一对特异性引物与TapMan探针,建立整套快速准确鉴定布鲁氏菌的实时定量荧光PCR检测体系。以实验室构建的克隆有布鲁氏菌目的片段的重组质粒为标准品,进行实时定量荧光PCR反应检测,通过优化反应条件,建立标准曲线。以标准品为模板,对该方法的特异性、敏感性与重复性进行检测与分析。并以临床样本进行检测的结果进行比较分析,结果表明,该检测体系的检测灵敏度高,重复性与特异性良好。本研究建立的实时定量荧光PCR可用于准确检测样本中的少量的布鲁氏菌,具有良好的应用前景和市场价值。     

动物布鲁氏菌病防治研究进展

布鲁氏菌病至今依然是在世界范围内流行的人畜共患传染病。除牛种、羊种、猪种、绵羊副睾种、犬种布鲁氏菌病的流行外，最近又发现了海洋种布鲁氏菌病的存在。对布鲁氏菌病的病原学诊断已从传统的分离培养鉴定发展到应用PCR、基因探针等分子生物学技术，诊断的敏感性大大提高了。在血清学诊断方面，EIISA技术已经成为国际公认的成熟技术。在预防用疫苗方面，牛用RB51和羊用Mlll两种粗糙型疫苗开辟了布鲁氏菌病防治的新领域，将更有效地推动布鲁氏菌病的防治效果。     

羊布鲁菌快速检测技术应用研究

通过用4种方法对羊布鲁菌的检测,确立适用于基层应用的羊布鲁菌快速检测技术。http://www.mdxycn.com/采用浊度仪LAMP、金属浴LAMP、荧光目视检测试剂盒LAMP和普通PCR共4种方法对羊血液中的布鲁菌进行检测,比较其检测结果。结果表明,共检测162份羊血液,布鲁菌检出率分别为浊度仪LAMP为8.6%、金属浴LAMP为8.6%、荧光目视检测试剂盒为8.6%、普通PCR为6.2%。LAMP较普通PCR方法特异性强且灵敏度高;金属浴LAMP方法操作简便,无需特殊设备,更适用于基层应用。