无乳链球菌Sip-GAPDH嵌合核酸疫苗的制备及免疫效果评价     

王蓓  李桂欢  鲁义善  汤菊芬  吴灶和  简纪常 《水产学报》2014,38(11):1926-1936

为了研究无乳链球菌Sip-GAPDH嵌合核酸疫苗对罗非鱼链球菌病的免疫保护作用,本研究通过PCR和重叠延伸拼接技术(SOEing)获得融合基因Sip-GAPDH,连接至真核表达载体pcDNA3.1(+)制备DNA疫苗,通过背鳍肌肉注射方式免疫吉富罗非鱼,免疫后第7、28天取样,采用PCR及RT-PCR方法检测pcDNA-Sip-GAPDH在各个组织(包括肌肉、脑、头肾、脾脏、鳃和肝脏)中的表达情况。采用ELISA、Real-time PCR法和体外攻毒法分别分析各实验组不同时间血清抗体效价、免疫基因(IgM、IL-1β和CD8)表达变化规律和相对保护率,研究该嵌合性疫苗对吉富罗非鱼的保护效果。结果显示,实验组在免疫接种第7和第28天时,注射点周围的肌肉、脑、头肾、脾脏、鳃和肝脏均能检测到嵌合性质粒,实验组罗非鱼血清抗体效价均高于对照组并于21 d时达到峰值(1∶4 096);qPCR数据表明,实验组鱼体胸腺、头肾和脾脏的IgM、IL-1β和CD8基因的mRNA表达量均出现了上调,并且在胸腺、头肾和脾脏中的表达量分别在免疫后第12和第48 h达到峰值;免疫后42 d进行人工攻毒后计算免疫保护率为93.3%。以上研究结果表明,本研究构建的无乳链球菌Sip-GAPDH嵌合核酸疫苗在罗非鱼链球菌病防治中具有潜在的应用价值。  相似文献   

Surface-exposed expression of Edwardsiella tarda EseB in live attenuated Vibrio anguillarum based on novel surface display systems     

Qiyao Wang  Zhao Yang  Qin Liu  & Yuanxing Zhang 《Aquaculture Research》2009,40(13):1459-1467

Live, attenuated Vibrio anguillarum strains can serve as vectors for the delivery of heterologous antigens for development of multivalent recombinant vaccines. Based on the outer membrane anchoring elements of V. anguillarum , we have previously constructed several efficient surface display systems Lpp-Omporf1, Lpp-OmpU, Lpp-Omp26La, Wza-Omporf1, Wza-OmpU and Wza-Omp26La. In this study, with these constructed surface display systems, a putative antigen protein EseB from pathogenic Edwardsiella tarda was successfully expressed on the surface of an attenuated V. anguillarum strain to get multivalent vaccine candidates. Further immune protection evaluation in zebra fish ( Danio rerio ) demonstrated that the V. anguillarum EseB-display strain AV/pW-26La-B could trigger full protection against V. anguillarum infection and early protection against E. tarda infection in the immunized fish. These results suggest that surface display of heterologous protective antigens in attenuated V. anguillarum could be used as a tool to develop potential V. anguillarum vector vaccine.  相似文献   

蜂胶佐剂对异育银鲫嗜水气单胞菌灭活疫苗的免疫增强试验   总被引：2，自引：0，他引：2  

肖琳琳  张庆华 《海洋渔业》2004,26(4):295-299

本文就蜂胶佐剂对鱼类细菌性疫苗的免疫增强作用进行了首次评价。试验以异育银鲫嗜水气单胞菌灭活疫苗为对象,比较了蜂胶佐剂、铝胶佐剂和无佐剂三种疫苗对银鲫的免疫效果。经抗体监测表明,免疫接种后第2周,蜂胶佐剂组银鲫已检测到血清抗体,随后抗体效价迅速上升,第4周达到峰值2^5：铝胶佐剂组和无佐剂组第3周检测到抗体．铝胶佐剂组第5周达到峰值2^5,无佐剂组第4周达到峰值2^4。银鲫免疫后第5周进行攻毒保护试验,发现蜂胶佐剂组、铝胶佐剂组和无佐剂组的免疫保护率分别为77．3％．65．0％和56．3％。试验结果表明,蜂胶佐剂能有效增强异育银鲫细菌性灭活疫苗的免疫保护力。  相似文献   

二种佐剂对哈氏弧菌OMPC-溶藻弧菌LPS偶联疫苗的增效作用     

李华  钱永华  石存斌  潘厚军  吴淑勤 《水产科学》2007,26(9):489-493

制备哈氏弧菌SpGY020601外膜蛋白复合物(OMPC)和溶藻弧菌EpGS021001脂多糖(LPS)偶联疫苗,设置偶联疫苗组、偶联疫苗 ISA763佐剂组、偶联疫苗 β-葡聚糖佐剂(DEAE)组以及生理盐水对照组,并以相同程序免疫卵型鲳鲹。溶菌酶活性检测、微量凝集反应试验结果显示:各疫苗组血清中的溶菌酶含量无显著差异(P>0.05),但均远高于生理盐水对照组(P<0.05);ISA763佐剂组抗哈氏弧菌SpGY020601抗体滴度较其他组出现的早,抗溶藻弧菌EpGS021001抗体滴度较其他组持续时间长;β-葡聚糖佐剂组次之,其次是无佐剂组,对照组始终未检测到抗体。各组分为腹腔注射攻毒与浸泡攻毒,对EpGS021001的攻击,ISA763佐剂组相对保护率分别为90%和70%,高于β-葡聚糖佐剂组的85%和55%,以及无佐剂组的80%和55%;对SpGY020601的攻击,ISA763佐剂组相对保护率分别为95%和80%,高于或相当于β-葡聚糖佐剂组的95%和75%,以及无佐剂组的90%和75%。  相似文献   

仔猪大肠杆菌病三价蜂胶灭活疫苗猪体血清抗体消长规律     

范忠军  柴虹  朱万光  李士成 《吉林畜牧兽医》2007,28(8):11-12,15

采用大肠杆菌C83549(O149:K88ac)、C83644(O68:K99)、C83710(O9:987P)菌株制备抗原,以蜂胶为佐剂,研制仔猪大肠杆菌病三价蜂胶灭活疫苗,疫苗成品检验均符合相关要求。将疫苗免疫妊娠母猪并用凝集试验测定其所产仔猪的抗体消长规律,并研究疫苗免疫保护相关性。结果表明:妊娠母猪产前40d和15d各免疫1次所产仔猪母源抗体效价较高,仔猪在整个哺乳期都可以得到保护。同时也可运用产前30d和10d进行免疫接种方法。  相似文献   

Genotyping of Bovine Viral Diarrhoea Viruses Isolated from Cattle in Northern Italy     

Falcone E  Cordioli P  Sala G  Tarantino M  Tollis M 《Veterinary research communications》2001,25(2):161-167

Following the first official report of a clinically severe outbreak of bovine viral diarrhoea disease occurring in a farm in northern Italy, which had originated from the use of a live vaccine contaminated with a strain of BVD genotype II virus, a retrospective study on the prevalence of BVDV genotypes in Italy became highly relevant. For this purpose, the genotype of 78 BVDV-positive specimens, obtained in 1998–1999 from dairy cattle in an area near to where the outbreak occurred, was characterized by PCR technology. Two sets of primers, spanning the 5 UTR of BVDV genome, were used sequentially in a first round of RT-PCR, performed on viral RNA extracted directly from 15 clinical samples and 63 BVDV-infected cell-culture fluids; a second PCR assay followed to selectively amplify only BVDV genotype II. All the viruses under study were characterized as BVDV genotype I. As well as contributing to a better understanding of the prevalence of BVDV genotypes in the field, the results of the present study illustrate the possibility that novel BVDV strains can emerge in susceptible animals through the use of contaminated immunobiological products for bovine use.  相似文献   

Two new variants of the Rhodococcus equi virulence plasmid, 90 kb type III and type IV, recovered from a foal in Japan     

Takai S  Murata N  Kudo R  Narematsu N  Kakuda T  Sasaki Y  Tsubaki S 《Veterinary microbiology》2001,80(4):373-381

Calves were vaccinated orally, subcutaneously or intraperitoneally with a smooth, plasmid-cured strain of Salmonella enterica serovar typhimurium, strain 81. Oral vaccination was not effective, as only 1/5 calves survived challenge with virulent S. typhimurium. Strain 81 was attenuated for calves, as only a slight rise in rectal temperatures was detected after vaccination. The organism was excreted by some calves in the faeces, but no signs of diarrhoea were observed after vaccination. After parenteral vaccination, strain 81 was able to reach the intestines, gastric associated lymphoid tissues and other internal lymphoid tissues and remained viable for up to 14 days in the bovine host. After oral challenge with a virulent strain, 9/10 vaccinated calves survived challenge as opposed to 4/10 control calves (p<0.5). Diarrhoea was present in all calves of the control groups, but in only 4/10 of the vaccinated calves. The clinical reactions of the vaccinated calves were milder than in the control calves, as the rises in rectal temperatures were lower, diarrhoea was less severe, and the challenge strain was present in fewer organs from vaccinated calves than control calves. This study showed that parenterally administered Salmonella vaccines can induce both mucosal and systemic immunity, and it is postulated that this capability of strain 81 is related to its colonisation of lymphoid tissues and other systemic and intestinal tissues. This study confirmed that plasmid-cured strains were attenuated in the bovine host and conferred significant protection after parenteral vaccination, but not oral vaccination.  相似文献   

“黄白散”对兔接种RHD疫苗后免疫功能的影响     

王新  邵红  朱战波  丁宝川 《畜牧与兽医》2001,33(4):8-10

在注射兔病毒性出血症 (RHD)疫苗后 ,30只兔随机分为 2组 ,试验组兔饲喂含 1 5 %黄白散的饲料。从接种到第 1 80天 ,每组取兔 8只 ,每隔 1 5d采血 1次 ,检测血凝抑制 (HI)抗体 ;在接种前 1天及接种后 5、 1 0、 1 7、 2 4、 31、 41、 5 1d ,每组取兔 1 0只 ,采血检测ANAE+淋巴细胞百分率 ;在 1 80、 2 4 0、 30 0d时 ,每组分别取兔 5只做攻毒保护试验。结果表明 :2组兔HI抗体峰值分别为 7 3log2和 9 8log2 ,差异极显著 (P <0 0 1 ) ;试验组疫苗保护期可延长 60d ;试验组ANAE+率从第 1 0天至 5 1天与对照组比较 ,差异极显著(P<0 0 1 )。所以黄白散可增强兔接种RHD疫苗后的特异性免疫功能 ,增强RHD疫苗的免疫效果。  相似文献   

油乳剂疫苗中硬脂酸铝含量与油相水相比例关系研究   总被引：2，自引：0，他引：2  

裴春生  何玉友  李群  唐艳琳  王玉莲 《中国兽药杂志》2005,39(2):21-23

进行了鸡新城疫、传染性支气管炎、产蛋下降综合征三联油乳剂疫苗生产中降低油相比例的试验.当油乳剂疫苗的油相中硬脂酸铝含量分别为2%、1%、0.5%,对应的油相:水相分别为3:1、2:1和1:1时进行的乳化试验结果表明:随着油相中硬脂酸铝含量降低,油相和水相的比例相应降低,呈正比例关系.当油相中硬脂酸铝含量与油/水比分别为2%和3:1、1%和2:1、0.5%和1:1时,制备的油苗质量均达到了国家标准.  相似文献   

转基因植物疫苗研究进展     

傅玲琳  帅江冰  许梓荣 《中国兽药杂志》2005,39(5):22-26

众多研究表明,病毒的抗原决定簇及细菌毒素亚单位均能在转基因植物中成功表达,并保持良好的免疫原性.与现有的疫苗生产体系相比,植物生产疫苗具有安全、经济、稳定、高效等优势.本文概述了利用不同受体植物生产疫苗的研究现状、免疫原性评价及免疫原基因的优化表达策略.  相似文献