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猪圆环病毒Ⅱ型特异抗体检测间接ELISA法的建立 总被引：1，自引：0，他引：1
利用巴斯德毕赤酵母表达的猪圆环病毒2型株Cap蛋白为包被抗原,成功地建立了一种检测血清中PCV-2特异抗体的间接ELISA方法.确定了血清样品阴、阳性判定标准.临床检测结果表明,自建ELISA试剂盒具有较高的敏感性和特异性. 相似文献
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利用鸡鲍氏志贺氏菌和鸡痢疾志贺氏菌制备多价凝集抗原,确定其最佳反应浓度及比例.选择合适的试验鸡只,用2种鸡志贺氏菌灭活疫苗定期肌肉注射免疫和抗体效价测定,适时采血分离血清.结果显示,鸡鲍氏抗原与痢疾抗原的最佳混合浓度为6×10~9mL~(-1),最佳比例为1:1.制备的鸡鲍氏和鸡痢疾志贺氏菌多价阳性血清,经抗体效价测定均达6log2,阴性血清与鸡志贺氏菌多价凝集抗原作用无凝集现象出现.研制的多价凝集抗原及其标准血清经特异性试验、重复性试验和保存方法试验,具有特异性强、敏感性高、重复性好、易保存等特点,且可省去单价凝集抗原繁琐的检测程序和时间. 相似文献
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Rabbits immunized with freshly isolated homologous thyroid microsomes in complete Freund's adjuvant developed a severe chronic thyroiditis with the concomitant production of antimicrosomal antibodies. The antibody response was measured by indirect immunofluorescence and complement fixation, employing isolated microsomes as the antigen. Microsomes but not thyroglobulin absorbed out the immunologic activity of the rabbit serum. 相似文献
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斑点叉尾鮰对3种致病菌灭活菌苗的免疫应答 总被引：1，自引：1，他引：0
将经福尔马林灭活的柱状嗜纤维菌(Cytophaga columnaris)、嗜水气单胞菌(Aeromonas hydrophi-la)和叉尾鮰爱德华菌(Edwardsiella ictaluri)作为免疫原,分别接种斑点叉尾鮰(Ictalurus punctatusRafinsque)后,通过测定受免鱼的交叉凝集抗体效价、头肾和血液中吞噬细胞的吞噬活性及用A.hydrophila活菌攻毒的方法,探讨了斑点叉尾鮰对3种灭活菌苗的免疫应答状况。试验结果表明,对斑点叉尾鮰经腹腔注射接种3种灭活菌苗能刺激受免鱼产生较强的免疫应答,受免鱼产生了对3种致病菌的特异性凝集抗体和交叉凝集抗体,与未接种菌苗的对照鱼相比,受免鱼头肾和血液中吞噬细胞的吞噬活性明显上升,活菌攻毒的结果也证明了受免斑点叉尾鮰对C.columnaris,A.hydrophila和E.ictaluri产生了不同程度的相对免疫保护率(RPS)。 相似文献
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用温酚法分别从柱状嗜纤维菌(Cytophaga columnaris)、嗜水气单胞菌(Aeromonas hydrophila)和叉尾鮰爱德华菌(Edwardsiella ictaluri)3种鱼类致病菌中提取粗脂多糖(LPS)作为免疫原,分别接种斑点叉尾鮰(Ictalurus punctatusRafinsque)后,通过测定受免鱼的交叉凝集抗体效价、头肾和血液中吞噬细胞的吞噬活性以及对A.hydrophila活菌细胞内的杀菌活性,探讨了3种致病菌的粗LPS对斑点叉尾鮰的免疫原性。结果表明,从3种致病菌中提取的粗LPS对斑点叉尾鮰均具有较强的免疫原性,受免鱼的血清中存在对3种致病菌的交叉凝集抗体,与对照鱼相比,头肾和血液中吞噬细胞对吞噬原的吞噬活性和对A.hy-drophila活菌杀菌活性明显上升。说明在3种供试菌的粗LPS上不同程度地存在3种致病菌的共同保护性抗原。 相似文献
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比较了3种苦马豆素(Swainsonine,SW)人工抗原对小鼠的免疫原性,筛选出最佳的蛋白载体,为制备疯草毒素疫苗奠定基础。活性酯法制备SW-OVA、SW-BSA、SW-HSA 3种SW人工抗原,冷冻干燥后,紫外分光光度法测定偶联率。将56只昆系小白鼠随机分为SW-OVA大剂量组(A组)、SW-OVA小剂量组(B组)、SW-BSA大剂量组(C组)、SW-BSA小剂量组(D组)、SW-HSA大剂量组(E组)、SW-HSA小剂量组(F组)和空白对照组(G组),每组8只。A~F组分别接种3种SW人工抗原,首次免疫抗原的大、小剂量分别为50μg/只和100μg/只,第15天进行第2次免疫,抗原量均为第1次免疫抗原量的1.5倍,第29天进行第3次免疫,抗原量均为第2次免疫抗原量的1.5倍,G组对照,注射等量生理盐水。第28天开始检测血清中SW抗体效价。紫外光谱测定结果显示,本试验制备的SW-OVA、SW-BSA、SW-HSA 3种SW人工抗原偶联率分别为13、10和19。ELISA测定结果显示,A和B组的SW抗体效价一直保持很高的水平,A和B组小鼠血清的SW抗体效价于第98天分别到达最高峰13(log2)和12(log2);C~F组的SW抗体效价较低。结果证实,SW-OVA作为理想的免疫原,能够刺激小鼠的免疫应答,诱导产生很高的SW抗体,在防治动物疯草中毒方面具有理想的应用前景。 相似文献
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Specific antigen in serum of patients with colon carcinoma 总被引：35，自引：0，他引：35
The binding of monoclonal antibody specific for colon carcinoma was inhibited by serum from patients with adenocarcinoma of the colon but not by serum from patients with other bowel diseases or from healthy volunteers. Of other malignancies studied, serum from two patients with gastric carcinoma and two patients with pancreatic carcinoma also inhibited the specific binding of monoclonal antibody. The levels of carcinoembryonic antigen in these serum samples were not correlated with their levels of binding inhibition. Such monoclonal antibodies may prove useful for the detection of colorectal carcinoma. 相似文献
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根据新城疲病毒(NDV)对鸡红细胞的凝集以及相应抗体对病毒血凝的抑制的原理设计出新城疫抗体监测试剂盒,主要用于鸡新城疫抗体水平的监测,为正确实施新城疫疫苗适时接种提供依据。试剂盒包括新城疫阳性血清、新城疲阴性血清、4单位新城疫抗原和10ml/L醛化红细胞。经对50多份血清检测结果表明,用于血凝(HA)和血凝抑制(HI)试验的醛化红细胞与新鲜红细胞无显著性差异(P>0.05),试剂盒能准确检测出鸡群新城疫抗体水平,具有操作简便,适用,易于保存等特点。 相似文献
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猪繁殖与呼吸综合征病毒重组N蛋白间接ELISA检测方法的建立 总被引：1，自引：0，他引：1
在最佳条件下,通过对重组质粒pGEX-6P-N进行诱导表达,并利用Glutathione Sepharose 4B亲和吸附柱对表达产物（重组N蛋白）进行纯化。用重组N蛋白作为包被抗原,通过对相关条件进行优化（如抗原包被量,血清稀释度,酶标二抗最佳工作浓度,底物作用时间等）,确定判定标准,最终初步建立了检测PRRSV抗体的间接ELISA方法。用建立的间接ELISA方法检测360份血清样品,并与IDEXX公司研制的ELISA试剂盒检测的结果相比较,批内重复性试验变异系数均值为1.52%,批间变异系数为6.17%,符合率达92.5%,试验表明,建立的间接ELISA检测方法具有良好的重复性和特异性,可用于PRRSV血清抗体的检测。 相似文献
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绵羊肺炎支原体间接血凝诊断方法的建立 总被引：3，自引：1，他引：2
用绵羊肺炎支原体抗原致敏经戊二醛处理的绵羊红细胞,制备成间接血凝试验抗原,通过IHA来检测绵羊支原体性肺炎血清抗体.致敏绵羊红细胞的最佳抗原浓度为100μg/mL,血清效价≥1∶8即为阳性,效价≤1∶4者判为阴性.该方法具有很好的敏感性、特异性和可重复性,而且操作简单,适合大范围推广应用. 相似文献
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Rabbit antibody with a sedimentation coefficient of 7S was digested with pepsin to yield the 5S fragment. Washed specific precipitates of 5S antibody and egg albumin, when added to guinea pig serum, fixed up to 40 percent of the total complement. This fixation could not be attributed to contamination with 7S antibody. Absorption of guinea pig serum with washed precipitates of 5S antibody and antigen removed that portion of complement which was fixed by fresh 5S antibody-antigen aggregates, but not that fixed by 7S antibody-antigen aggregates. The presence of 0.001M ethylenediaminetetraacetic acid prevented this absorption of complement by the 5S aggregates. 相似文献
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Specific suppression of immune responses 总被引：17，自引：0，他引：17
D A Rowley F W Fitch F P Stuart H K?hler H Cosenza 《Science (New York, N.Y.)》1973,181(105):1133-1141
The models we have discussed in detail demonstrate specific suppression of immune reactivity produced in normal adult animals by antibody and antigen. The mechanism of homeostasis of suppression in these models depends on continued exposure to antigen and on an active response by the host. The active response may include production of antibody directed against specific receptors as well as antibody directed against antigen. Thus, specific regulation of both antibody and cell mediated immunity to an antigen might be achieved by the use of only the biological agents of the response: antigen, antibody, and possibly antibody to receptors. The general implication is that these same biological agents are responsible for autoregulation of immune reactions occurring in nature. Presumably, these agents may be used to suppress or reverse immune responses for appropriate clinical objectives. 相似文献
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A specific precipitate of ovalbumin and its rabbit-serum antibody, after fixing human serum complement, was digested with papain. The digest was analyzed immunochemically for complexes of antigen, antibody fragments, and components of complement. The results indicated that complement is not bound to Porter fragment III, but very likely is bound to fragments I and II. 相似文献
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Antigen competition: a paradox 总被引：7，自引：0，他引：7
R H Waterston 《Science (New York, N.Y.)》1970,170(962):1108-1110
Immunization of mice with pig erythrocytes caused impairment of the antibody response to subsequent immunization with sheep erythrocytes, a phenomenon called "antigen competition." Paradoxically, spleen cells from mice previously injected with pig erythrocytes produced an increased response when immunized in vitro with sheep erythrocytes. Augmentation of the in vitro response is due to an increase in one of the interacting cell types. "Antigen competition" is not due to competition for cells. Cell transfer experiments provided evidence that "antigen competition" observed in animals is the result of a humoral factor, presumably antibody, present in the animal but eliminated during preparation of cells for culture. 相似文献
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目的；制备抗磷酸酪氨酸抗体。方法：以EDC（碳化二亚胺）交联法合成磷酸酪氨酸牛血清白蛋白（P－Tyr－BSA）；以P－Tyr－BSA免疫新西兰免，抗血清经硫酸该盐析和DEAE纤维素柱层析纯化；以琼脂糖双扩法测定抗体效价。结果：得到抗血清效价为1：32，抗血清与牛血清蛋白（BSA）交叉反应的效价为1：2。纯化的抗体与磷酸酪氨酸牛血清白蛋白、磷酸酪氨酸卵清蛋白、磷酸丝氨酸牛血清白蛋白、磷酸苏氨酸牛血清白蛋白反应效价分别为1：32、1：16、1：2、1：2，所得抗体蛋白量为120mg。纯化的抗体在醋酸纤维素薄膜上电泳为单一区带。结论：用此法制备得的抗体专一性高，交叉反应低，抗体量大。 相似文献
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斑点叉尾鮰对3种致病菌外膜蛋白(OMP)的免疫应答 总被引：1，自引：1，他引：0
利用从嗜水气单胞菌(Aeromonas hydrophila)、柱状嗜纤维菌(Cytophaga columnaris)和叉尾鮰爱德华菌(Edwardsiella ictaluri)中提取的外膜蛋白(outer membrane protein,OMP)作为免疫原,注射接种斑点叉尾鮰(Ictalurus punctatus Rafinsque)后,通过测定受免鱼的交叉凝集抗体效价、肾脏和血液中吞噬细胞的吞噬活性和采用A. hydrophila活菌攻毒方法,比较了3种致病菌OMP对斑点叉尾鮰的免疫原性。试验结果表明:从3种致病菌中提取的OMP对斑点叉尾鮰均具有较强的免疫原性,受免鱼的血清中存在对3种致病菌的交叉凝集抗体,与对照鱼相比,肾脏和血液中吞噬细胞对吞噬原的吞噬活性和对A. hydrophila活菌攻毒的相对免疫保护率(relative percent survival,RPS)明显上升。说明在3种供试菌的OMP上不同程度地存在共同保护性抗原。 相似文献
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A rapid procedure for purification of rabbit antibody to beta-D-galacto-sylphenylazo-bovine serum albumin is described. Antibody was precipitated with a heterologous antigen and was then dissociated from the precipitated complex with hapten. The antigen was partially removed by pH adjustment, and antibody was separated from residual antigen and from hapten by partition chromatography on Sephadex G-25. The antibody, obtained in 50 to 55 percent over-all yield, was 96 percent reactive with homologous antigen. It appeared to consist, of 95 percent of a single component, based upon physicochemical measurements. 相似文献
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小鹅瘟羊源高免血清的制备及比较 总被引：1，自引：0，他引：1
为制备高效价的小鹅瘟高免血清,试验采用从贵州某养鹩场病鹅分离的小鹅瘟野生强毒株,接种于9 d鹅胚,收集24 h以后死亡的鹅胚尿囊液,制备小鹅瘟病毒抗原,与弗氏佐剂混合均匀,多次免疫山革后,颈动脉一次性放血,离心收集血清即得到小鹅瘟羊源高免血清,以琼脂扩散进行血清效价测定,并与市售小鹅瘟鹅源高免血清进行保护率比较.结果显示,试验制备的小鹅瘟羊源高免血清抗体效价在1:8以王,对人工感染雏鹅的保护率为95%,而市售小鹅瘟鹅源高免血清的保护率只有85%.结果表明,本试验制备的小鹅瘟羊源高免血清具有效价高、成本低、安全性好等优点. 相似文献