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为制备驴源抗犬细小病毒(canine parvovirus,CPV)抗体,首先利用F81猫肾细胞扩增CPV-2a病毒株,经甲醛灭活和氢氧化铝胶乳化,制成CPV灭活疫苗,病毒滴度为10-8.5 TICD50,然后通过肌肉分点注射免疫驴,连续免疫3次(第1次注射3mL,后2次各注射6mL),每次免疫前采血,分析驴血清的抗体滴度和中和抗体效价;利用盐析法和离子交换层析法分离和纯化驴血清IgG。用IgG处理幼犬(50mg/kg),用CPV-2a毒株对IgG处理的犬进行攻毒,分析IgG抗CPV的活性。结果显示,免疫3次后驴血清的CPV抗体滴度为1∶8 192,中和抗体效价为28.5。用制备的IgG处理幼犬未发现不良反应,经CPV-2a攻毒实验证实IgG处理犬的发病率和死亡率分别为33%,0%,均明显低于非处理组(分别为100%,83%),说明制备的驴源IgG具有明显的抗CPV活性。 相似文献
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鹅卵黄IgG的纯化及兔抗鹅IgG酶标抗体的制备 总被引：4，自引：0，他引：4
采用三氯甲烷去脂、无水Na2S04盐析、DEAE23纤维素层析相结合的方法纯化鹅卵黄IgG,SDS-PAGE检测表明,所得鹅卵黄IgG纯度可达93%;用纯化的IgG免疫家兔,抗血清经双向琼脂扩散,证明兔抗鹅IgG抗血清效价约1:64,免疫电泳试验检测,证明得到了特异性抗血清,提取兔抗血清的IgG,用辣根过氧化物酶标记,制备了兔抗鹅IgG的酶标抗体,酶标抗体的效价为1:6400. 相似文献
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用自制IgG型兔抗猪IgG、IgA、IgM型抗体,建立BA-ELISA检测乳清和血清中IgG、IgA、IgM型抗LT抗体的方法,经临床应用表明,BA-ELISA方法不仅灵敏度高、重复性好,而且可分别检测IgG、IgA、IgM型抗体,该法可作为评价能表达LT的大肠杆菌菌苗免疫效果和研究大肠杆菌免疫机理的方法之一。 相似文献
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为了制备兔抗鸽IgG抗体-HRP,建立斑点酶联免疫吸附试验定性检测鸽毛滴虫抗原的检测方法,将纯化的鸽IgG加入佐剂免疫兔子,获取纯化的兔抗鸽IgG抗体;采用简易过碘酸钠法标记兔抗鸽IgG,获取兔抗鸽IgG抗体-HRP;同时将鸽毛滴虫用超声波粉碎加入佐剂后多次免疫健康鸽获取鸽抗鸽毛滴虫高免血清(一抗);然后在2×106个/mL的鸽毛滴虫抗原量下,用不同浓度的一抗抗体、兔抗鸽IgG抗体-HRP进行试验;并对70个临床样品进行检测。结果显示,在2×106个/mL的鸽毛滴虫抗原量下,一抗与兔抗鸽IgG-HRP最佳工作浓度分别为1∶100和1∶500;70个样品中,48个镜检为阳性的样品用Dot-ELISA检测有44个呈阳性,阳性符合率为91.7%;22个镜检为阴性的样品用Dot-ELISA检测有12个呈阴性,阴性符合率为54.5%;Dot-ELISA检测与镜检结果的总符合率为80%。结果表明,本试验成功地建立了鸽毛滴虫感染的Dot-ELISA检测方法。 相似文献
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应用猪繁殖与呼吸综合征病毒HuB-20株活疫苗免疫兔,制备兔抗猪繁殖与呼吸综合征病毒毒株的特异性抗体IgG,应用15%SDS-PAGE和Western-blot进行纯化鉴定.结果表明:抗猪繁殖与呼吸综合征病毒HuB-20株特异性抗体IgG经15%SDS-PAGE纯化分析和Western-blot的鉴定显示该特异性抗体IgG具有很好的免疫原性,特异性强,仅与该病毒毒株的ORF5发生特异性结合.说明兔抗猪繁殖与呼吸综合征病毒HuB-20株特异性抗体具有良好的免疫原性和特异性,可与猪繁殖与呼吸综合征病毒ORF5特异性结合,可以作为诊断试剂和诊断试剂盒的一抗. 相似文献
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间接酶联免疫吸附测定法(ELISA)的第二抗体,根据国内外文献记载,必须与第一抗体相对应。如检测患马血清的抗体,必须制备兔抗马IgG;检测患牛血清的抗体,必须制备兔抗牛IgG。从1980年以来,我们用ELISA检测牛伊氏锥虫病抗体,发现第二抗体可用免抗马IgG;用ELISA检测马伊氏锥虫病抗体,第二抗体亦可用兔抗牛IgG,现将结果报道如下。 相似文献
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de Freitas JC Lopes-Neto BE de Abreu CR Coura-Vital W Braga SL Reis AB Nunes-Pinheiro DC 《Research in veterinary science》2012,93(2):705-709
This research investigated the profile of anti-Leishmania antibodies in different clinical forms of canine visceral leishmaniasis (CVL). Naturally infected dogs were divided into two groups: subclinical dogs (SD, n=10) and clinical dogs (CD, n=68). Non-infected dogs (ND, n=7) comprised the negative control group. The humoral response was evaluated by the profile of total IgG, IgG1, IgG2, IgM, IgA and IgE, determined by ELISA. Infected animals showed increased levels of total IgG, IgA and IgE in addition to IgG1 and IgG2 in groups SD and CD, when compared with group ND. Furthermore, it was observed that IgG2 and IgM were correlated with symptomatology, while total IgG, IgG1 and IgA were negatively correlated and IgE showed no correlation. It follows that serum levels of IgG2 anti-Leishmania are correlated with typical clinical signs of disease. Furthermore the determination of specific anti-Leishmania antibodies could be an important tool in monitoring CVL clinical picture. 相似文献
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The comparative opsonic efficiency of ovine salmonella-specific antibody isotypes was determined by measurement of specific phagocytic uptake of opsonised virulent Salmonella typhimurium by ovine mammary neutrophils. An in vitro phagocytosis assay revealed that IgM was superior to IgG2 in promoting the phagocytosis of opsonised virulent organisms. IgG1, on the other hand, was non-opsonic. Superiority of the IgM isotype over IgG2 as an opsonin was also evident in studies on the viability of opsonised S typhimurium upon phagocytosis. It was revealed that the percentage of organisms killed was appreciably greater when opsonisation was carried out with the IgM than with the IgG2 isotype, although after ingestion by neutrophils there was essentially no difference in the efficiency with which the ingested organisms were killed. 相似文献
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M T Jensen L J Kemeny S S Stone A E Ritchie 《American journal of veterinary research》1980,41(1):136-139
Porcine colostral immunoglobulin (Ig)G and IgA, isolated from transmissible gastroenteritis virus-infected sows, were compared by direct immunoelectron microscopy. It was estimated, using antibodies with a less than a twofold difference in virus-neutralizing activity, that IgG was 500 times more efficient than was IgA for coating transmissible gastroenteritis virions. Guinea pig complement enhanced the antibody coating with IgG, but did not increase virus-neutralizing activity of IgG or IgA. 相似文献
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Labelled swine albumin resp. IgG were injected intravenously in two boars. The radioactivity of seminal plasma was determined at frequent intervals. It was observed that minimal amounts of radioactivity of albumin and IgG could be detected in seminal plasma. The importance of a transmission of IgG from serum to seminal plasma is discussed. 相似文献
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Gnotobiotic calves were inoculated by the intratracheal route with Mycoplasma bovis and the specific antibody response in sera and tracheo-bronchial washings examined by radioimmunoassay. In sera an IgM response which reached a peak two weeks post infection was followed by IgG1 and IgG2 antibody responses. Low levels of IgA antibody were detected in sera three and four weeks after infection. The predominant antibody in tracheo-bronchial washings 2 weeks after infection was IgA. Four weeks after infection IgG1 antibody predominated, but IgG2 and IgA antibodies were also present. Cells containing Ig were present in the cellular accumulations around the necrotic zones produced by M. bovis in the lung parenchyma two and four weeks after infection. IgG1 containing cells predominated in these cellular infiltrates. IgG2 producing cells were the next in frequency. It is concluded that the lung lesion caused by M. bovis is partly due to the host's immune response, presumably contributing to the control of the infection, and that the cells infiltrating the lung are a major source of the local and systemic IgG antibody that is detected after infection. IgA staining cells were observed in the submucosa of tissues from nasal cavity and trachea. These cells are probably the source of IgA in tracheo-bronchial washings and sera since IgA-producing cells were not a predominant component of the lesion in the lung parenchyma. 相似文献
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《Journal of Equine Veterinary Science》1998,18(11):742-748
Newborn foals, deprived of colostrum and its rich supply of immunoglobulin G (IgG), were supplemented both orally and intravenously with purified equine immunoglobulin G (Lyphomune®). Data were obtained from 18 foals given oral administration of IgG at Colorado State University and 26 foals given IgG intravenously at the Jockey Club de Sao Paulo in Brazil.Oral administration of 10-gm doses of Lyphomune® in 18 colostrum-deprived Arabian foals, at various intervals within the first 24 hours after birth, resulted in increased serum concentrations of IgG. Administration of one 10-gm dose of Lyphomune® immediately following birth provided a mean serum IgG level of 125 mg/dl after two hours. The recommended dosage of two 10-gm doses per 15 kg of body weight produced mean IgG serum concentrations of approximately 400 mg/dl by 14 hours. It was determined that an early bolus of IgG was most effective, although administration at any period during the first 24 hours would increase IgG levels significantly and in direct relationship to grams of Lyphomune® administered.After the 24-hour study period, colostrum from each respective mare was provided by bottle feeding (200 ml) to 10 of the foals that were then allowed to nurse their dams normally. Significant increases in circulating IgG were observed in nine of these ten animals at four and eight hours after colostrum administration. No interfering effect was noted when colostrum and Lyphomune® were given to the same foal.Intravenous administration of 10-gm doses of Lyphomune® in Thoroughbred foals, immediately after birth, resulted in serum concentrations of IgG of 200–300 mg/dl six hours later. A second intravenous dose, at six hours after the initial dose, resulted in an additional average increase of 184 mg/dl. Four of six foals administered 10 gm of Lyphomune® for each 15 kg of body weight reached serum concentrations greater than 400 mg/dl. It was demonstrated that Lyphomune® was able to increase circulating levels of IgG, by either oral or intravenous administration, to levels considered protective in the newborn foal. 相似文献
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T W Jungi T Francey M Brcic B Pohl E Peterhans 《Veterinary immunology and immunopathology》1992,33(4):321-337
Ovine bone marrow-derived macrophages (BMM) may express several IgG receptor (Fc gamma receptor; FcR) subsets. To study this, model particles (opsonized erythrocytes; EA), which are selectively handled by certain FcR subsets of human macrophages were used in cross-inhibition studies and found to react in a similar manner with FcR subsets of sheep macrophages. In experiments with monoclonal antibodies against subsets of human FcR, human erythrocytes (E) treated with human anti-D-IgG (anti-D-EAhu) and sheep E treated with bovine IgG1 (Bo1-EAs) were handled selectively by human macrophage FcRI and FcRII, respectively. Rabbit-IgG-coated sheep E (Rb-EAs) were recognized by FcRI, FcRII and possibly also by FcRIII of human macrophages. Anti-D-EAhu, Bo1-EAs and Rb-EAs were also ingested by sheep BMM. Competitive inhibition tests, using various homologous and heterologous IgG isotypes as fluid phase inhibitors and the particles used as FcR-specific tools in man (anti-D-EAhu and Bo1-EAs), revealed a heterogeneity of FcR also in sheep BMM. Thus, ingestion of anti-D-EAhu by ovine BMM was inhibited by low concentrations of competitor IgG from rabbit or man in the fluid phase, but not at all by bovine IgG1, whereas ingestion of Bo1-EAs was inhibited by bovine IgG1. This suggested that anti-D-EAhu were recognized by a FcR subset distinct from that recognizing bovine-IgG1. It was concluded that sheep BMM express functional analogs of human macrophage FcRI and FcRII and that Bo1-EAs and anti-D-EAhu are handled by distinct subsets of BMM FcR. All EAhu tested (EAhu treated with anti-D, sheep IgG1 or sheep IgG2) were ingested to a lower degree than EAs. This inefficient phagocytosis could be enhanced by treatment of EAhu with antiglobulin from the rabbit, suggesting that it is caused by a low degree of activity of opsonizing antibodies rather than special properties of the erythrocytes themselves. Several lines of evidence suggested that both FcR subsets of ovine BMM recognize both ovine IgG1 and IgG2. In contrast, bovine IgG1 reacts with one FcR subset and bovine IgG2 interacts inefficiently with all FcR of ovine BMM. 相似文献
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L Rodák 《Veterinary immunology and immunopathology》1984,5(4):377-387
Class-specific antibodies against bovine IgG1, IgG2, IgM and IgA and porcine IgG, IgM and IgA immunoglobulins were prepared. Their class specificity was assessed by two radioimmunological methods, namely, radioimmunoelectrophoresis and double antibody sandwich radioimmunoassay. The methods are highly specific and sensitive and do not require the use of purified immunoglobulins, but can be performed with normal serum or colostrum. It was confirmed that antibodies found satisfactory in these tests were suitable for a wide range of use including radioimmunoassay and enzyme linked immunosorbent assay. 相似文献
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E Leresche L Paunier G Andrejevic 《Comparative immunology, microbiology and infectious diseases》1979,1(3):205-220
The aim of this study was to investigate the stability of bovine immunoglobuline and maintenance of their efficacity given per os to newborns, and the tolerance to this preparation. This preparation is designed for prevention and/or treatment of infant gastroenteritis by passive immunity.It contains bovine immunoglobulines IgG (75% of total), IgM and IgA with a high titer against frequent serotypes of E. coli. It was given per os to 10 newborns, 8 being aged 1 to 5 months, and 2 aged 18 months. Samples of stool were collected before (controls) and after treatment (tests). Bovine IgG were estimated in the stools by standard immunodiffusion test, specific titer against coli antigens by agglutination technic. Blood formula was checked throughout.Tolerance to the preparation was good. In the stools of 9 of the 10 newborns, bovine immunoglobulines were found at higher concentrations in tests than in controls, both in term of quantity (several mg IgG per gram of stool) and in term of specificity against E. coli serotypes. The stools of one child only (aged 18 months) was negative for bovine IgG and specific immunoglobulines.These results demonstrate that bovine immunoglobulines of this preparation given per os are stable through the gastrointestinal tractus and that their specificity is maintained, in particular in children aged less than 5 months. 相似文献
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Isotype-specific ELISAs for the detection of antibodies to bovine respiratory syncytial virus 总被引：1，自引：0，他引：1
T G Kimman F Westenbrink P J Straver D Van Zaane B E Schreuder 《Research in veterinary science》1987,43(2):180-187
Isotype-specific ELISAs for the detection of antibodies to bovine respiratory syncytial virus (BRSV) are described. BRSV-specific IgG1 and IgG2 were determined in indirect double antibody sandwich assays. For IgA and IgM antibody capture assays were used. The isotype specificity of the assays was confirmed by the observation that samples with a high titre of BRSV-specific antibodies of particular isotype were negative in the assays for the other isotypes and vice versa. Comparison of the results obtained in the ELISAs and in the virus neutralisation test showed that acute phase antibodies were more efficiently detected in the latter. It also showed that the presence of BRSV-specific IgA was not correlated with neutralising activity in vitro. The serum antibody response of BRSV-infected seronegative calves from the field consisted of a nearly simultaneous increase of IgM, IgA and IgG1-antibodies in the acute phase of the disease, while the IgG2-response followed at various intervals thereafter. In young animals with maternal antibodies a different pattern was found. There was no increase in IgG1 and IgG2, but six of eight animals showed a weak IgM response and two of these six calves also showed a weak and short lasting IgA response. Because maternal antibodies are insufficiently effective in protecting calves against BRSV, the presence of such antibodies at mucosal surfaces was investigated. Maternal immunity was found to be restricted to IgG1 antibodies in serum. This agrees with the failure of maternal antibodies to protect mucosal surfaces against BRSV infection. 相似文献