猪圆环病毒2型抗体免疫金标检测试纸的研制

应用酶联免疫原理和胶体金层析技术,采用特殊的生产工艺,在玻璃纤维膜包被胶体金标记PCV-2抗原,在硝酸纤维素膜上检测线和对照线处分别包被PCV-2抗原和兔抗PCV-2抗体,制成猪圆环病毒2型抗体免疫金标检测试纸。当待检样品阳性时,在检测线处形成抗原抗体的免疫复合物而凝聚显色;当待检样品阴性时,检测线处不形成抗原抗体免疫复合物不显色。整个试验过程只需15min。试纸与ELISA试剂比较,两者都具有微量、特异、准确的优点,且金标试纸独具操作方便、快速和结果直观、容易判定的优点

铁硫二元团簇的产生及其紫外光解

用激光蒸发固体样品的方法产生了各种铁硫原子簇离子Fe_nS_m~+(n=1-13,m=1—13),并且用串级飞行时间质谱仪研究了这些团簇离子的紫外激光解离规律.实验发现,所有组成为m=n,m=n-1与m=n+5的离子均具有较大的丰度,表明它们具有较好的稳定性.各种Fe_nS_m~+离子的紫外光解结果显示出,当nn)离子的结构为在Fe_nS_n~+核心外围再键合上其他的S 原子

Study and Apply of Colloidal Gold Strip in Detecting the Antibody of pseudorabies

应用免疫学原理与胶体金层析技术相结合 ,采用双抗原夹心法建立了检测猪伪狂犬病抗体水平的“猪伪狂犬病抗体免疫胶体金快速检测试纸法” ,用该法与美国Idexx公司的猪伪狂犬病ELISA试剂盒 ,同时对 1 6份猪血液或血清进行抗体水平检测 ,符合率达 1 0 0 %。同时使用金标法对 1 0 0份猪血液和对应血清进行检测 ,结果也相同。试验结果显示 ,金标法与ELISA一样 ,具有微量、特异、准确等优点 ,且操作简易 ,而金标法不需要任何仪器 ,检测时间短 ,结果直观 ,容易判定 ,适用于基层兽医站、养猪场使用和大面积猪伪狂犬病抗体普查。Based on the principle of colloidal gold immunochromatography, a test strip was prepared and applied in detecting the antibody of pseudorabies. The test result was coincidence with ELISA made in USA Indexx cor. in examination of 80 blood or sera samples from pig、chicken、 duck、 cattle and goat. It shows that this strip has the same merits as that of ELISA test in microanalysis, high specialty and accurate. No special instrument is needed and the test can be completed easily in a short time. So it can be used widely in common veterinary services, hoggery and the general survey of pseudorabies disease

猪圆环病毒2型ORF1和ORF2基因的克隆与表达

根据GenBank猪圆环病毒2型基因序列,设计合成两对特异性引物,对厦门株-1进行PCR扩增,得到ORF1和ORF2基因。将PCR产物酶切后,插入到pET-22b载体中,构建原核表达载体pET-22b/ORF1、pET-22b/ORF2,转化大肠杆菌BL21(DE_3),经IPTG诱导表达,收集茵液进行SDS-PAGE分析。确定重组蛋白主要以包涵体的形式表达,包涵体洗涤溶解后,采用Ni~(2+)离子金属螯合亲和层析柱纯化蛋白,逐步透析法进行复性。Western blot和ELISA分析结果表明Rep蛋白和Cap蛋白均能与猪圆环病毒2型阳性血清发生特异性反应

Study on Colloidal Gold Strip in Detecting Classical Swine Fever Virus

应用胶体金标记技术(Immunogold Labelling Technique)与免疫层析技术相结合,在玻璃纤维膜、硝酸纤维膜的检测线(T)和对照线(C)上分别喷上胶体金与猪瘟(CSF)单克隆抗体(Ab2)的偶联物、CSF单克隆抗体(Ab2)和羊抗鼠IgG多克隆抗体,制成检测CSF抗原的"猪瘟抗原胶体金快速检测试纸"。用该试纸检测"猪瘟弱毒活疫苗"和猪瘟脾淋弱毒苗均显阳性,而对猪源性的其它病毒、细菌抗原均显阴性。经临床测试,该试纸具有微量、特异、快速、简便和结果容易判定的优点,可作为检测猪瘟抗原的一种新试剂。Based on the principle of immunogold labelling technique and immunochromatography,the coupling product from colloidal gold and CSF McAb2 was sprinkled on the glassfiber film; CSF McAb2 and polyclonal sheep anti-mouse IgG were sprinkled on the detection line and control line of nitrocellulose membrane,colloidal gold strip was executed to detect CSF antigen quickly.The results were positive using the strips to detect the CSF antigens.To detect other avian viruses appeared negtive with strips.The colloidal gold immunochromatography technology is a new,visual,driblet,specific,rapid and simply detection method,and it is easy to adjust the results