树突状细胞在沙门氏菌感染获得性免疫中的作用

树突状细胞（DC）在沙门氏菌入侵部位存在,并具有从上皮细胞组织向次级淋巴器官迁移的能力,加之具有激活幼稚型T细胞的功能,DC的这些特性决定了它们在启动针对沙门氏菌感染的获得性免疫应答的过程中将发挥关键作用.本文就DC在沙门氏感染获得性免疫中的作用作一综述.     

口服鼠疫F1-V重组融合基因DNA活菌疫苗的构建及免疫原性研究

目的 构建携带鼠疫耶尔森菌F1-V融合基因的重组减毒沙门菌苗,口服免疫Balb/c小鼠检测其免疫原性,为口服鼠疫活载体DNA疫苗研究打下基础.方法 将F1-V融合基因克隆到真核表达载体asd-pVAX1,进一步依次将重组质粒转化减毒沙门菌X3730、X4550得到重组沙门菌X4550(asd-pVAX1/F1-V),提取重组质粒转染COS-7细胞并做免疫组化和Western-blot检测F1-V融合蛋白在细胞中的表达.以1×109CFU/只的剂量3次口服免疫Balb/c小鼠,ELISA方法检测血清中抗体水平.结果 构建的重组减毒沙门菌转染COS-7细胞后,免疫组化和Western-blot试验证明F1-V融合蛋白在细胞中得到了瞬时表达,ELISA检测到免疫小鼠血清有特异性抗体IgG产生.结论 构建的重组沙门菌能运送DNA疫苗到体内并成功释放质粒刺激机体产生特异性免疫应答,为口服鼠疫活载体DNA疫苗的黏膜免疫研究打下了基础.     

沙门氏菌鞭毛蛋白属共同抗原表位的鉴定

应用针对沙门氏菌鞭毛蛋一具有广谱反应性的单抗证实了该蛋白上存在共同抗原表位，进一步鉴定了鞭蛋白基因表达产物，结果同样表明该类共同抗原表位的存在。通过对该共同抗原表位在沙门氏菌属内外的分布规律测定，显示出它的属特异性。用单抗ＣＢＢ和display status     

应用PCR技术快速检测市售猪肉中产单核李斯特菌

目的：调查安徽省产单核李斯特菌对动物性食品的污染状况。方法：采集合肥农贸市场生猪肉样品，通过李斯特菌富集培养基培养16h，应用PCR技术进行产单核李斯特菌的检测。结果：从120份猪肉样品中分离出2株产单核李斯特菌，阳性检出率为1．7％。结论：表明安徽省市售猪肉中有不同程度产单核李斯特菌的污染，且存在李斯特菌病的可能性。整个检测过程只需24h，显示PCR技术对产单核李斯特菌的检测具有特异性强、灵敏度高、速度快和易操作等优点。     

空肠弯曲菌生物膜研究进展

空肠弯曲菌是全球范围内主要的人兽共患性、细菌性肠道病原菌之一[1],其感染率在世界各地普遍呈上升趋势,主要引起人体急性肠炎和食物中毒,并伴发反应性关节炎、肝炎、瑞特氏病和格林-巴利综合征等免疫性损伤性疾病[2]。     

合肥市屠宰生猪主要微生物学指标调查研究

目的：了解合肥市定点屠宰场生猪屠宰加工产品的卫生质量以及加工生产的卫生状况。方法：应用国标法对5个定点生猪屠宰场500份生猪胴体体表样品及200份胴体肉样进行菌落总数、大肠菌群和沙门菌的检测。结果：生猪胴体肉样菌落总数和大肠菌群的总超标率达46%和22.5%;生猪胴体体表样品和肉样沙门菌检出率分别为24.2%和17%;大肠菌群和沙门菌之间呈现正相关。结论：合肥市生猪胴体的卫生质量急需提高,屠宰生产加工水平亟待改善,在肉品生产过程中重视大肠菌群的控制则有利于降低沙门菌的污染程度,实施宰前管理和宰前检验是保证病健隔离分宰,减轻对加工环境和产品污染的重要环节。     

甲型H1N1流感病毒HA基因的原核表达及免疫反应性分析

目的构建甲型H1N1流感病毒HA基因的原核表达质粒,获得融合表达蛋白,并对表达蛋白的免疫反应性进行分析。方法人工合成A/California/05/2009 H1N1流感病毒的HA基因,以合成基因为模板,通过PCR方法扩增出去除信号肽的HA部分基因片段,然后将去除信号肽的HA基因克隆至pET30a(+)原核表达载体中,构建出原核表达质粒,再将重组质粒转化E.coliBL21(DE3)表达菌株;重组菌经IPTG诱导后,收集菌体进行SDS-PAGE电泳分析,Western blot分析表达产物的免疫反应性。结果获得了HA基因的原核表达重组菌,细菌经IPTG诱导表达后,SDS-PAGE分析可见约67.4 ku大小的目的蛋白表达条带,Western blot结果显示,表达产物与人甲型H1N1流感患者阳性血清具有反应性。结论成功表达出甲型H1N1流感病毒的HA蛋白,该蛋白具有良好的免疫反应性,为甲型H1N1流感的快速诊断方法的建立提供了生物材料。     

宠物重要人兽共患细菌病流行现状及其控制策略

随着我国社会经济发展和城市化进程的加速,宠物数量日益增多,宠物人兽共患病发病率日渐增高,本文对我国宠物重要人兽共患细菌病流行状况、原因进行了阐述,并提出了防控对策.     

重组沙门菌表达结核分枝杆菌ESAT-6及其特异性免疫应答分析

Objective To determine the immune responses induced by recombinant Salmonella ty-phimurium expressing the secreting antigen ESAT-6 of Mycobacterium tuberculosis. Methods ESAT-6 cod-ing gene was cloned and identified by PCR and sequencing. Prokaryotic expression plasmid pYA33-esat car-rying the ESAT-6 coding sequence was constructed firstly and electro-transformed into an attenuated strain X4550 of Salmonella typhimurium, the recombinant bacteria was named as X4550(33-esat). C57BL/6 mice were immunized intranasally (I. N) with 108 CFU recombinant bacteria at day 0 and 18. Cells from spleen, lung, mesenteric lymph node (MLN) and Peyer's patch (PP) were collected from mice after second immu-nization, and the specific IFN-γ-secreting cells and IL-4-secreting cells were detected by ELISPOT assay u-sing ESAT-6 peptide as stimulus. Furthermore, CTL effects were in vivo evaluated by CFSE assay. Results The results showed that cellular immune responses specific for ESAT-6 could be detected by ELISPOT assay. In lung and PP cells, immune responses against ESAT-6 were biased toward Th1 type, the frequency of IFN-γ-secreting cells was much higher than that of IL-4-secreting cells. In splenocytes and MLN cells, the anti-gen specific immune responses acted as Thl and Th2 balance, the frequency of IFN-γ-secreting cells was close to that of IL-4-secreting cells. CFSE assay indicated that recombinant bacteria could induce the high level of CTL effects specific for ESAT-6 peptide. Conclusion These results suggested that recombinant Sal-monella typhimurium X4550(33-esat) not only can induce cellular immune responses, but also can elicit specific CTL responses after I. N immunization. It also provided the useful information for the control of infec-tious disease of tuberculosis.