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检测羊感染的捻转血矛线虫（Ｈａｅｍｏｎｃｈｕｓｃｏｎｔｏｒｔｕｓ，Ｈｃ）的两种酶免疫分析方法的比较（摘要）Ｓｃｈａｌｌｉｎｇ，Ｈ．Ｄ．等我们以成熟的Ｈｃ排泄或分泌的产物（ＥＳ）和虫体粗提物（ＣＳＡ）作为抗原建立的两种酶免疫分析方法，比较他们分别检测抗... 相似文献
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对１０２例临床型和１８７例隐性型乳房炎病例的奶样进行了细菌学分析。临床型病例中，８５．３％的乳样检出了细菌，检出率高于隐性乳房炎病例（７２．２％）。传染性病原，包括无乳链球菌（Ｓｔｒｅｐｔｏｃｏｃｃｕｓａｇａｌａｃｔｉａｅ），金黄色葡萄球菌（Ｓｔａｐｈｙｌｏｃｏｃｃｕｓａｕｒｅｕｓ）和牛棒状杆菌（Ｃｏｒｙｎｅｂａｃｔｅｒｉｕｍｂｏｖｉｓ），在临床型病例中的检出率是４１．２％，隐性型病例中的检出率是３９．５％，是检出率最高的一类致病菌。临床型乳房炎的另一类重要致病菌是环境性细菌。本研究检出的这类致病菌有乳房链球菌（Ｓｔｒｅｐｔｏｃｏｃｃｕｓｕｂｅｒｉｓ），粪链球菌（Ｓｔｒｅｐｔｏｃｏｃｃｕｓｆａｅｃａｌｉｓ），牛链球菌（Ｓｔｒｅｐｔｏｃｏｃｃｕｓｂｏｖｉｓ），埃希氏大肠杆菌（Ｅｓｃｈｅｒｉｃｈｉａｃｏｌｉ）和变形杆菌（Ｐｒｏｔｅｕｓｓｐｐ．），检出率是２０．６％，而隐性乳房炎中的检出率只有２．２％。凝固酶阴性的葡萄球菌（Ｃｏａｇｕｌａｓｅ－ｎｅｇａｔｉｖｅｓｔａｐｈｙｌｏｃｏｃｃｉ，ＣＮＳ）在两类乳房炎中的检出率分别是１４．７％和１３．４％。此外还检出了一些乳区有两种或两种以上的细菌混合感染。 相似文献
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日本血吸虫谷胱甘肽S—转移酶小鼠免疫试验 总被引：3，自引：0，他引：3
用谷胱甘肽琼脂糖亲和层析柱纯化中国大陆株日本血吸虫成虫谷胱甘肽Ｓ－转移酶（ＧＳＴ）。把纯化的ＧＳＴ结合福氏佐剂免疫了二批ＢＡＬＢ／ｃ小鼠，获得了２９．５８～３２．７１％的减虫率和５２．９４～６８．１３％，粪便减卵率，ＥＬＩＳＡ试验表明免疫小鼠产生了体液免疫应答，免疫印迹试验（Ｗｅｓｔｒｎ Ｂｌｏｔｔｉｎｇ）显示日本血吸虫水溶性成虫抗原和ＧＳＴ抗原的２８、２８ｋＤ蛋白被免疫小鼠血清所识别。 相似文献
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对猪生殖与呼吸综合征病毒（ＰＲＲＳＶ）核衣壳蛋白（Ｎ）作为ＥＬＩＳＡ抗原进行了研究，将编码ＰＲＲＳＶＮ蛋白的基因０ＲＦ７ｃＤＮＡ插入杆状病毒表达载体ｐＢｌｕｅＢａｃＨｉｓ２Ａ，通过同源重组获得了重组杆状病毒ＯＲＦ７－ＡｃＭＮＰＶ，感染昆虫细胞ＳＦ９表达了Ｎ蛋白，占细胞总蛋白的７．０７％，纯化后作为抗原建立了间接ＥＬＩＳＡ，与ＩＤＥＸＸ公司的ＥＬＩＳＡ诊断试剂盒有９６％的符合率，与ＩＦＡ有１００％的符合率。试验证实：ＰＲＲＳＶＮ蛋白在昆虫细胞中得到高效表达且是检测ＰＲＲＳＶ抗体的良好抗原。 相似文献
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以大鹅观草（ＲｏｅｇｎｅｒｉａｇｒａｎｄｉｓＫｅｎｇｅｔ．Ｓ．Ｌ．Ｃｈｅｎ）与竖立鹅观草（Ｒ．ｃｉｌｉａｒｉｓｖａｒ．ｊａｐｏｎｅｎｓｉｓ（ｈｏｎｄａ）Ｂ．Ｒ．Ｌｕ，Ｙｅｎｅｔ．ｙａｎｇ２ｎ＝２８，ＳＳＹＹ）犬草（Ｅｌｙｍｕｓｃａｎｉｎｕｓ（Ｌ．）Ｌ，２ｎ＝２８，ＳＳＨＨ）拟鹅观草（Ｐｓｅｕｄｏｒｏｅｇｎｅｒｉａｓｐｉｃａｔａ（Ｐｕｒｓｈ）Ａ．ｌｏｖｅ，２ｎ＝２８，ＳＳＳＳ）三个种进行杂交，对亲本 相似文献
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猪细小病毒生物学特性研究 总被引：2，自引：0，他引：2
应用ＰＫ－１５传代细胞复制猪细小病毒（ＰＰＶ）参考株ＮＡＤＬ－２和云南分离株ＫＭ，研究得出细胞培养物半数细胞感染量分别为１０＾－７．９／ｍｌ和１０＾－６．８／ｍｌ经ＣｓＣｌ密度梯度离心，两种病毒颗粒其浮力密度值分别为１．３３ｇ／ｃｍ＾３和１．３９／ｃｍ，电镜观察病毒粒子呈圆形，直径约２０ｎｍ。ＳＤＳ－聚丙烯酰胺凝胶电泳（ＳＤＳ－ＰＡＧＥ）分析，表明纯化的ＰＰＶ细胞培养毒，主要含两种蛋白成分，分子量 相似文献
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鸡传染性喉气管炎ＥＬＩＳＡ诊断方法研究──ＳＰＡ─ＥＬＩＳＡ检测ＩＬＴＶ抗原方法的初步建立彭万强，黄承锋，吕敏娜，孔庆飞，朱治远，幸桂香，杨海金（广东省农科院兽医研究所广州．５１０６４０）葡萄球菌Ａ蛋白（ＳａｐｈｙｌｏｃｏｃｃｕｓＰｒｏｔｅｉｎＡ─Ｓ... 相似文献
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应用SDS—PAGE和IEF电泳对伊氏锥虫和媾疫锥虫的蛋白组分进行了分析.在SDS—PAGE中,伊氏锥虫有21条带.媾疫锥虫有19条带,两种虫体的蛋白质区带在分子量40000～90000之间存在差异,尤其表现在表面糖蛋白上,伊氏锥虫为43000,媾疫锥虫为60000.在IEF电泳中,伊氏锥虫出现26条带,媾疫锥虫出现33条带,两种虫体的蛋白质区带在等电点4.8～5.6之间相同,而在5.6～7.0之间存在差异.本实验还对马媾疫锥虫的蛋白质进行了双相电泳分析,显示出86个多肽斑点. 相似文献
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Thin-layer agarose isoelectric focusing: an improved technique for determining sheep hemoglobin type
An improved technique for rapid screening of sheep flocks for hemoglobin (Hb) type is presented. This technique, isoelectric focusing (IEF) on thin-layer agarose gels is simple, rapid, inexpensive and is suitable for screening large numbers of sheep for Hb type. With this technique, up to 100 sheep blood samples can be prepared, tested and interpreted within 2 h after samples are drawn. The new technique was shown to provide better resolution than polyacrylamide gel electrophoresis (PAGE) and was able to resolve samples in which the Hb had become partially degraded. These same samples could not be resolved by PAGE. The use of a special electroendosmosis-free grade of agarose provided resolution essentially equal to polyacrylamide as a matrix for IEF. The advantages are that the casting of the agarose gels is considerably easier, the focusing of samples is more rapid, staining and destaining times are greatly reduced and hazards from potential neurotoxicity of acrylamide are eliminated. Blood from 138 ewes at the Oregon State University Sheep Center was examined by the new agarose IEF technique to determine and demonstrate its usefulness for screening. No difficulty was encountered with interpretation of any of the samples. Frequencies of the HbA and B alleles were similar to those found in earlier studies when polyacrylamide tube gel electrophoresis was used. The observed frequencies were also similar to those expected with the population in Hardy-Weinberg equilibrium. 相似文献
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为建立高致病性猪繁殖与呼吸综合征病毒(HP-PRRSV)蛋白质组双向电泳方法,本研究通过对HP-PRRSV蛋白样品提取、裂解、一向等电聚程序、上样量、染色试剂、显色时间等条件优化,建立了有效的HP-PRRSV蛋白质组学双向电泳方法。优化结果表明采用冻融-超声-裂解提取样品,结合2-D clean-up试剂盒纯化蛋白,按上样量为200μg,采用硝酸银染色,显色6 min,选择适宜的一向等电聚焦参数,能获得分辨率高、重复性好的双向电泳图谱。HP-PRRSV蛋白质组双向电泳方法的建立,为开展该病毒蛋白质组和免疫蛋白质组研究奠定了基础。 相似文献
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用几种电泳方法分析牦牛肝片吸虫不同部位抗原 总被引：1，自引：0，他引：1
应用ＳＤＳ－ＰＡＧＥ、等电聚焦电泳（ＩＥＦ）及双向电泳三种电泳方法分析牦牛肝片吸虫成虫四种抗原（头抗原－ＨＡ、体抗原－ＢＡ、体表抗原ＳＡ、分泌排泄抗原－ＥＳ）。ＳＤＳ－ＰＡＧＥ结果表明，ＢＡ、ＨＡ、ＳＡ分子量在１２～１００ｋＤ之间，ＥＳ在１２．３～２６ｋＤ之间，蛋白质染色ＢＡ、ＨＡ、ＳＡ、ＥＳ分别显示２５、２４、１８、９条带；ＩＥＦ分析肝片吸虫分别得３４、３３、２８、２２条带，主要由酸性蛋白质组成，主要谱带在ｐＩ４．２０～６．５５内；双向电泳分析ＢＡ、ＥＳ多肽斑点为６９、３０个 相似文献
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C Salomoni M Fiorentino D L Palenzona E Di Maio G De Benedictis 《Comparative immunology, microbiology and infectious diseases》1987,10(2):125-132
The effects of some environmental and genetic factors on the isoelectric focusing (IEF) patterns of muscle proteins from a fish Lebistes reticulatus (Cyprinodonts) were investigated. Of two diets applied the restricted one resulted in a loss of bands in the IEF patterns of the treated fishes muscle homogenate. Instead immunization gave an increase in both number and intensity of IEF bands in (4.0-6.0; 7.0-7.9) pH ranges which are typical of immunoglobulins. Lastly IEF patterns from random and assortative mating animals were compared, resulting in loss of IEF-bands in the inbred lines. Results showed that IEF of muscle proteins could be an excellent method to evaluate the response by fish phenotypes to environmental or genetic factors. 相似文献
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桑突变体Cty-Ym叶片蛋白质的双向电泳及质谱分析 总被引：2，自引：0，他引：2
为探讨桑树叶色突变体CtyYm变异的分子机理,采用蛋白质组学分析技术,从突变体和对照桑中提取并分离叶片蛋白质,获得了IEF/SDSPAGE双向电泳图谱,在CBB染色条件下,检测到比较清晰的蛋白质点184个;经PDQuest软件分析,发现两者蛋白质点的相对含量有36处存在显著差异(±2倍),尤其是突变体中蛋白质点SSP2801的相对含量仅为对照桑的258%,经胶内酶切、LCESIQTOFMS分析、肽质量和碎片离子检索分析,鉴定该蛋白组分为RuBisCO大亚基,推测突变体CtyYm叶色黄化可能与RuBisCO大亚基含量减少有关。 相似文献
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Cyst fluid antigens of Echinococcus granulosus, Taenia hydatigena and T pisiformis were examined by electrophoresis using homologous and heterologous hyperimmune rabbit sera to these antigens. While arc 5 forming antibodies were identified in sera from rabbits immunised with E granulosus and T hydatigena cyst fluids, antibodies responsible for forming precipitating antigen B band were detected in rabbit antisera to E granulosus, T hydatigena and T pisiformis antigens. T hydatigena cyst fluid appears to contain antigen similar to E granulosus antigen 5 and probably antigen B while T pisiformis cyst fluid has mainly an antigen close to hydatid antigen B. 相似文献
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Díaz F Massó F Páez A Varela E Suaréz-Güemes F Montaño LF 《Veterinary immunology and immunopathology》1999,67(3):203-212
Due to the complexity and variety of biological effects found in Mycobacterium bovis (M. bovis) proteins analyzed solely on a molecular weight (MW) basis, we approached the purification of M. bovis proteins through their isoelectric point (pI). Twenty M. bovis culture filtrate protein extract (CFPE) isoelectric focused (IEF) protein fractions, confined between pI3 and 10, were isolated. The MW of the major proteins isolated in the various fractions correlated with protein already reported 14-, 18-, 20-, 25-, 31-, 38-, 45-, 64-, 67- and 70 kDa by SDS-PAGE. Since several different pI fractions showed proteins of the same MW we tested the ability of all IEF fractions to stimulate interferon-gamma (IFN-gamma) production by peripheral blood mononuclear cells (PBMC) isolated from cattle with well defined M. bovis tuberculosis (TB) infection. In animals with few lesions IFN-gamma inductive IEF fractions were in the acid range. As the number of lesions increased, neutral fractions were also inductive. Some fractions with relatively few proteins induced as much IFN-gamma production as others with abundant proteins. None of the 20 IEF fractions enhanced IFN-gamma production by anergic cells. We conclude that IFN-gamma production in diseased animals is induced mainly by acidic mycobacterial proteins and that the response towards these proteins is enhanced as the disease progresses, what coincides with higher PPD reactivity. However, the IFN-gamma production in anergic status was severely affected. We found that this cytokine production is spontaneous and antigen-independent. 相似文献