注射用布氏菌活疫苗免疫学评价

目的以小鼠为动物模型,通过检测布氏菌活疫苗免疫小鼠产生的体液免疫、细胞免疫应答及保护力,对其进行免疫学评价。方法将30只小鼠随机分为3组,皮下免疫布氏活疫苗,免疫4周后分离血清及脾脏淋巴细胞。ELISA法检测免疫动物血清中总IgG抗体,抗体亚类IgG1及IgG2a;ELISPOT法检测分泌IFN-γ、IL-4的脾脏淋巴细胞数目,以及用ELISA方法检测体外再刺激后小鼠脾细胞分泌IFN-γ、IL-4的细胞因子水平;用羊布氏菌弱毒株M5攻击免疫动物,通过脾脏细菌计数评价布氏活疫苗的免疫保护作用。结果用ELISA法检测到免疫布氏活苗的小鼠体内有特异性抗体产生;ELISPOT可检测到较高的分泌IFN-γ脾脏淋巴细胞数目,ELISA方法可检测到较高的IFN-γ水平,表明布氏活疫苗主要诱导Th1型免疫应答;通过布氏活疫苗保护力分析,表明该疫苗具有较好的免疫保护效果。结论布氏疫苗采用注射免疫途径是可行的,免疫后能获得较好的免疫效果。     

布鲁氏菌血培养阳性报警时间的分析

目的:探讨布鲁氏菌血培养阳性报警时间的特点及临床意义.方法:选择2017年1月~2018年12月124例布病患者为观察组,随机选择性别、年龄相仿的其它细菌感染者为对照组,其中葡萄球菌112例,肠杆菌科菌91例.回顾性分析各组血培养的阳性报警时间.结果:布鲁氏菌48h内未见阳性报警者,48~72h、72~96h、96~120h、120~168h的阳性报警率分别为1.6%、14.5%、45.2%、35.5%;葡萄球菌和肠杆菌科菌48h内阳性报警率分别为87.5%、94.5%,均未见超过120h阳性报警者.组血培养平均阳性报警时间分别为:布鲁氏菌117.4h(103.4h^126.8h)、葡萄球菌24h(18h^34h)、肠杆菌科菌11h(9.5h^15h).组间比较差异有统计学意义(P<0.01).结论:布鲁氏菌血培养阳性报警时间明显长于于其它病原菌,对疑似布病的患者,血培养的孵育时间至少设置为7d.综合分析血培养阳性报警时间与典型的菌落、革兰氏染色形态,对鉴定布鲁氏菌具有重要意义.     

The Brucellae and Their Success as Pathogens

Brucellae are tiny, aerobic, slow growing, catalase and oxidase positive Gram negative coccobacilli or small rods, which may reach man through exposure to tissues of mammalian hosts via cuts or aerosols, or as food infections mostly through dairy products. As parasites brucellae are extraordinarily successful, causing very long-lasting infections in all mammalian social animals, such as ungulates, canids, and rodents; recently they have been found to also cause disease in pinnipeds and cetaceans. Brucellae as members of the alpha Proteobacteria, have suffered major losses of genomic material as they adapted to their facultative intracellular parasite role, and are able to initiate infection with minimal disturbance of the innate immune system, thus reaching a privileged intracellular niche where they multiply. Brucellae are likely to be among the toughest organisms to control through public health and agricultural policies, even involving detection-slaughter strategies.     

Renal Cyst Infection Caused by Brucella abortus

A 49-year-old man with a medical history of polycystic kidney disease was presented to the emergency department with fever and left flank pain. Abdominal examination revealed an enlarged and painful left kidney. The C-reactive protein level was significantly high and the magnetic resonance imaging revealed areas of abnormal intensity and fluid–fluid levels in renal cysts. Brucella abortus was yielded from both blood and cyst fluid culture. Standard therapy (rifampicin plus doxycycline) of brucellosis was started, but the clinical and laboratory signs subsided after the addition of ciprofloxacin. There was no need for aspiration of infected cyst fluid. Hereby, according to the medical database search, we report that the first renal cyst infection caused by B. abortus was successfully treated with triple antibiotic therapy.     

布鲁氏菌PFGE分子分型的条件优化

[目的]将PFGE方法应用于布鲁氏菌分型研究,探索建立布鲁氏菌PFGE分型的最优化方法。[方法]在用PFGE方法进行布鲁氏菌基因分型的研究中,针对不同内切酶、酶切的浓度和时间、脉冲时间等影响PFGE分型结果的因素进行优化,利用优化后的条件对布鲁氏菌19株标准菌株进行分型研究。[结果]在布鲁氏菌的PFGE分型研究中,选择XbaI为内切酶、酶切浓度为96u/200μl、酶切时间为4h、脉冲时间为2～25s为PFGE电泳的优化条件,在分析的19株布鲁氏菌标准菌株中,6种布菌的PFGE图谱各不相同,但PFGE不能将种内不同生物型布菌完全区分开来。[结论]PFGE方法可用于布鲁氏菌的基因分型的研究,是对传统分型方法一种较好的补充。     

PCR产物直接测序快速识别病原菌

[目的]结合PCR和DNA测序技术,建立一种适用于传染性疾病预防控制的快速准确地识别病原细菌的分子检测方法.[方法]以布鲁氏菌为例,利用PCR分别从纯培养物、模拟标本中扩增布鲁氏菌16S和31kd基因,并对扩增产物测序,与数据库进行比对,根据比对结果确定扩增产物是否来源于布氏菌.[结果]从纯培养物和模拟标本成功扩增特异产物,测定序列与数据库比对,能很快准确确定病原的种类,优于单纯的PCR扩增.[结论]结合PCR扩增和产物测序的分子检测方法切实可行,为疾病预防控制体系中病原菌的快速识别提供了一个快速、高效和准确的方法.     

布鲁氏菌病疫情的计算机多媒体信息系统的研究

目的为达到布鲁氏菌病疫情的共享性、快捷性、方便性及准确性,同时可为布鲁氏菌病防治专业人员及有关领导对布鲁氏菌病综合防制及监测等进行决策提供科学依据。方法以Windows98/2000/NT作为平台,利用Microsoft Powerpoint进行研制。结果建立了中国、内蒙古及赤峰市布鲁氏菌病相关疫情的多媒体信息系统。结论布鲁氏菌病疫情计算机多媒体信息系统有利于科研、教学和防治指导。     

布鲁氏菌猪种标准株外膜蛋白OMP25的原核表达

目的克隆布鲁氏菌外膜蛋白Omp25基因并构建基因的原核表达系统。方法用聚合酶链反应技术扩增得到布鲁氏菌基因组，得Omp25基因片段，T—A克隆后测定核苷酸序列，将目的基因定向插入原核表达载体pGEX-4T-1，经双酶切和DNA测序测定，将构建的重组质粒转化大肠杆菌（E．coli HB101，TOP10），经IPTG诱导后，用SDS—PAGE检测重组蛋白rOrap25表达情况。结果经DNA测序显示Omp25DNA序列和插入位点正确，成功构建了重组质粒DGEX-4T-1-Omp25，经IPTG诱导，特异性表达出以包涵体形式存在48kDa的Omp25融合蛋白。结论制备、克隆了布鲁氏菌外膜蛋白Omp25基因片段，并在大肠杆菌中表达出Omp25融合蛋白。     

The Antibody Response to Brucella: Immunoglobulin Response Measured by Enzyme-linked lmmunosorbent Assay and Conventional Tests

Abstract: The antibody response to Brucella: immunoglobulin response measured by enzymelinked immunosorbent assay and conventional tests. G. L. Gilbert and L. A. Hawes, Aust. N.Z. J. Med ., 1981, 11, pp 40–45.
An enzyme-linked immunosorbent assay was adapted to measure total and individual classes of brucella antibody. The results were compared with those of conventional tests for brucella antibody on the sera of a number of healthy seropositive abattoir workers and several patients with either acute or suspected chronic brucellosis. IgG was the class of brucella specific immunoglobulin most commonly detected in all groups. IgM was present in the sera of 40% of seropositive abattoir workers, all but one of the patients with recent acute brucellosis or seroconversion and none of those with suspected chronic brucellosis. Many of the abattoir workers' sera which contained brucella specific IgM gave negative results in the direct agglutination test. The presence of brucella specific lgM in the sera of these men was, in most cases, associated with no past history of acute brucellosis and a relatively short period of employment in the abattoir. It is suggested that the presence of brucella specific IgM in the serum of a person occupationally exposed to 6. abortus, probably indicates a relatively recent primary infection, either symptomatic or sub clinical and has no prognostic significance. Repeated or prolonged exposure is associated with IgG brucella antibodies, often in high titre, irrespective of symptoms. It was not possible, on the basis of any serological tests performed in this study to distinguish healthy people exposed to brucella from those with symptoms consistent with chronic brucellosis