特异性IgY抗体对中性粒细胞吞噬功能的影响

鸡卵黄免疫球蛋白 (Yolk Immunoglobulins,Ig Y)是一种存在于鸡蛋黄中的 Ig G。Ig Y比哺乳动物Ig G多一个 CH4区 ,为 7S免疫球蛋白 ,分子量约为1 80 k Da,含 67～ 70 k Da的重链和 2 2～ 30 k Da的轻链 [1 ]。抗绿脓杆菌、葡萄球菌及沙门氏菌的特异性Ig Y抗体能够抑制绿脓杆菌生长 ,抑制葡萄球菌产生肠毒素以及抑制沙门氏杆菌结合人肠细胞 (Caco2 ) [2 ] 。抗绿脓杆菌 Ig Y含漱预防胆囊纤维化患者绿脓杆菌感染 [3]。虽然 Ig Y不激活补体 ,不与哺乳动物细胞的 Fc受体结合 ,但 Ig Y在抗细菌性感染方面能发挥抗菌和防止炎症发生的作…     

一致性检验比较两种弓形虫抗体检测试剂盒

比较两个弓形虫抗体检测试剂盒检测结果的一致性,为流行病学调查和临床诊断筛选可合理使用的商品化试剂盒。对510份猪血清分别进行弓形虫抗体IHA和ELISA检测,应用Kappa检验比较两种试剂盒检测结果的一致性并进行分析,用x~2检验分析差异性。两种试剂盒联合检出79份阳性,阳性符合率为67.52%,阴性符合率为95.17%,总符合率为88.82%。x~2检验差异显著(x~2=228.23,p=0.00);u检验值为15.89,两种产品的一致性为中度一致(Kappa值=0.66)。所以弓形虫病IHA检测试剂盒可作为现场收益流行病学调查,而弓形虫Ig G抗体ELISA检测试剂盒适合辅助临床检测和诊断。x~2检验比较两种试剂盒具有局限性。u检验排除偶然性造成的一致程度,与Kappa检验结合可比较一致性。弓形虫抗体检测要根据实况选择不同的试剂盒。     

羊胎盘转移因子康肽对猫免疫功能的影响

研究康肽(羊胎盘转移因子)对猫免疫功能的影响。将28只猫分为4组(即对照组,康肽低剂量组、中剂量组、高剂量组),灌胃不同剂量的康肽,于试验前、后对免疫球蛋白Ig G、Ig A、Ig M、补体C3、补体C4及WBC、LY、RBC进行测定比较,其中WBC、LY、RBC用自动血液细胞分析仪检测,Ig G、lg M、lg A和补体C3、补体C4分别用Ig G、lg M、lg A试剂盒和补体C3、补体C4试剂盒进行检测。结果表明,康肽能提高猫血清中免疫球蛋白(Ig G、Ig M、Ig A)、补体(C3、C4)和WBC、LY、RBC的含/数量。提示康肽具有正向调节作用,剂量越大,作用越好。     

卵黄抗体在水产动物疾病防治中的应用

<正>卵黄抗体又称卵黄免疫球蛋白(Ig Y),一般以鸡作为供体,经一种或多种特定抗原免疫诱导后在卵黄中蓄积,机体所产生的特异性抗体,具有类似哺乳动物Ig G的免疫活性,可有效防治特定疾病。近年来,在畜禽疾病的诊断与防治方面卵黄抗体的运用已非常广泛,可将其作为抗生素替代品或饲料添加剂等进行使用,在动物养殖业中具有广阔的应用前景。目前,用卵黄抗体防治水产动物疾病的研究报道较少,但因其具有疗效确切、使用安全、无毒副作用、制备工     

一颗耀眼的科技创新的新星——第四军医大学动物保健品研制中心新型产品连创佳绩

第四军医大学动物保健品研制中心利用四军大的高精尖设备与先进技术 ,发挥关中毛驴这一自然资源扰势 ,采用新型工艺和独特保护剂 ,认真实践与创新 ,研制出科技含量较高的科研新产品“犬用多联活疫苗”和精致浓缩型“犬、猫、猪、羊、兔、鸡、鸭、鹅、鸽、鸵鸟等 1 0多种动物用抗多病免疫球蛋白”,成果产品“犬五联活疫苗”已申请国家专利 ,专利号为 :95 1 0 6 744· 3。鸽用抗多病免疫球蛋白 ( Ig G)专利申请号为 :991 1 0 85 8· 2犬、猫用抗多病免疫球蛋白 ( Ig G)专利申请号为 :991 1 0 85 9· 0猪用抗多病免疫球蛋白 ( Ig G)专利申请…     

基于鸡毒支原体热休克蛋白DnaK的间接ELISA方法的评价及初步应用

为研究鸡毒支原体(MG)重组热休克蛋白DnaK (rDnaK)的免疫反应原性及其作为ELISA方法中诊断抗原的应用价值,本研究将已构建的含有MG DnaK基因的重组质粒p ET-30a-DnaK进行原核表达纯化,以多份MG的标准阳性血清和阴性血清作为一抗,进行western blot分析,结果显示r DnaK与MG的阳性血清发生特异性反应,与MG的阴性血清无反应条带,表明r DnaK具有较好的免疫反应原性。将其作为包被抗原包被酶标板,用HRP标记的兔抗鸡Ig G作为酶标二抗,分别对抗原包被浓度、血清稀释度、酶标二抗稀释度及工作时间等反应条件进行了优化,建立了一种稳定检测MG抗体的间接ELISA方法。对该方法进行评估,灵敏性试验结果显示该方法与进口试剂盒相当;特异性试验结果显示包被抗原r DnaK不与NDV、IBV、ILTV等几种常见的禽呼吸道疾病的阳性血清发生交叉反应;重复性试验结果显示,批内变异系数为1.6%～5.15%,批间变异系数为2.9%～5.98%;符合率试验结果显示该方法与进口试剂盒的总体符合率为90.4%;抗体持续期试验结果显示该方法最早能在鸡群免疫MG疫苗后一周检测到MG抗体,表明该诊断方法适用于MG感染的早期检测。采用该方法检测了来自全国5个省份的568份临床样品,其结果显示MG的感染率在23%～51%,对我国MG的流行状况做了初步的调查。     

从新城疫免疫鸡所产卵的卵黄液中提取IgG方法的研究

应用硫酸萄聚糖钠沉淀的方法从新城疫免疫鸡卵黄液中提取出高效价纯净的IgG,平均浓度为10.839mg/ml,每枚鸡卵可提取出162 mg IgG,其HI抗体效价与提纯前的无明显差异。提纯的卵黄Ig G在—30℃低温冰箱内保存1年、在冻干状态下保存1.5年,抗体活性仍无明显影响。     

鸡减蛋性病毒诊断试剂盒研制成功

近日 ,由天津市畜牧兽医研究所黄金海副研究员主持完成的“鸡减蛋性病毒诊断试剂盒的研制与应用”通过了天津市科委组织的专家验收。该试剂盒可一次诊断出4种病毒。该项目的研究揭示了鸡血清抗体与局部抗体的关系 ,其中血清抗体与卵黄抗体密切相关 ,与胆汁抗体水平有一定的相关性 ,但与气管IgA抗体水平差异较大。该项目研究建立了鸡减蛋性病毒病抗体检测EHSA方法 ,可用于血清、卵黄和气管局部分泌抗体的检测。在此基础上 ,研究人员应用抗鸡Ig轻链单克隆抗体酶结合物 ,研制出鸡新城疫、禽流感、传染性支气管炎、减蛋综合症4种病毒病抗体水…     

鸡胚血清中被动抗体吸收规律的研究

<正>研究以双抗体夹心ELISA法检测420枚(只)从鸡胚9日龄至雏鸡5日龄的血液抗体Ig G变化趋势,结果发现血清抗体Ig G从0.27μg/m L递增至18.27μg/m L,并以雏鸡3日龄时血液中Ig G抗体含量最高。1材料与方法1.1鸡Ig G的提取与鉴定Ig G从蛋鸡血清中提取,按何昭阳等[1]报道的方     

鸡传染性喉气管炎ELISA试剂盒的应用研究

通过对不同来源的鸡传染性喉气管炎病毒(ILTV)血清的检测,进行ELISA试剂盒检测ILTV抗体的应用研究,并与传统的琼脂扩散试验(AGP)方法进行了比较。研究结果表明该试剂盒具有较好的敏感性、特异性和重复性,与AGP方法相比,其敏感性大大提高。ILT诊断用ELISA试剂盒可以应用于养禽生产实践中。     

Characterization of monoclonal antibodies to bovine IgG1

Monoclonal antibodies were developed to bovine IgG1. In addition, production of monoclonal antibodies to bovine light chain is also reported. Monoclonal antibody specificities were initially determined by solid-phase enzyme immunoassay. The monoclonal antibovine IgG1 was shown by a specificity-independent isotyping solid-phase enzyme immunoassay to be mouse IgG1 with kappa light chains. Ascites derived monoclonal antibovine IgG1 antibodies were linked to cyanogen bromide-activated Sepharose and used for affinity isolation of bovine IgG1. The bovine IgG1 eluted from the affinity column was characterized using immuno-electrophoresis, acrylamide gel electrophoresis, isoelectric focusing and solid-phase enzyme immunoassay. Affinity chromatography using monoclonal antibodies provided both a verification of monoclonal antibody specificity and a rapid technique for the isolation of bovine IgG1. This technique may also be employed to remove IgG1 contaminants during purification of bovine IgA.     

Isolation and characterization of peacock Chlamydophila psittaci infection in China

The objective of this study was to isolate and identify suspected pathogens from peacocks and peacock farmers with severe pneumonia and to investigate its potential association with peacocks' pneumonia, caused by Chlamydophila psittaci infection. A clinical examination of infected peacocks identified birds with symptoms of anorexia, weight loss, yellowish droppings, airsacculitis, sinusitis, and conjunctivitis, whereas the infected farmers showed high fever and respiratory distress. Immunofluorescence tests detected chlamydial antigens in pharyngeal swabs (12 of 20) and lung tissue samples (four of five) from peacocks. One of four swabs taken from farmers was also positive by the same test. Specific anti-chlamydia immunoglobulin G was detected in 16 of 20 peacocks and four of four peacock farmers. The isolated pathogen was able to grow in specific-pathogen-free (SPF) chicken embryos and McCoy cell lines and was identified as Chlamydiae by immunofluorescence assay and PCR. Avian influenza virus, Newcastle disease virus, and infectious bronchitis virus were eliminated as potential causative agents after pharyngeal swabs inoculated onto the chorioallantoic membrane of embryonate eggs failed to recover viable virus. PCR and restriction fragment length polymorphism indicated the ompA gene from the isolate was similar to that of avian C. psittaci type B. Three-week-old SPF chickens challenged with the peacock isolate via intraperitoneal injection showed a typical pneumonia, airsacculitis, and splenitis. Subsequently, the inoculating strain was recovered from the lungs of challenged birds. This is the first report of C. psittaci infection in peacocks and peacock farmers.     

两种免疫酶试剂盒检测猪抗E型肝炎病毒IgG的比较

目的比较两种免疫酶试剂盒recomWell HEV IgG(swine)ELISA(recom Well )和recomLine HEV IgG(swine)(recom Line )检测猪抗E型肝炎病毒(hepatitis E virus,HEV)IgG的准确性。方法分别采用基因3型和基因4型人HEV(human HEV,hHEV)人工感染无菌猪各2头,采用上述试剂盒检测猪感染后不同天数的血清样品中抗HEV IgG,同时对两头接种PBS液的无菌猪血清样品18份和4头未感染猪的18份血清进行了检测。结果 2头猪人工感染基因3型hHEV后,recomWell检测均于感染后21d出现阳性,并持续56d;recomLine检测,其中1头猪于感染后14d呈阳性,而另1头于感染后21d呈阳性,且均持续56d。2头猪人工感染基因4型hHEV后,两种试剂盒检测,其中1头猪于感染后21d呈阳性,另1头35d呈阳性,并均持续56d仍为阳性。18份对照血清两种试剂盒检测均为阴性。18份未人工感染猪血清中,只有1份两种试剂盒检测均为阳性。两种试剂盒同时检测的共72份样品中,recomWell检出阳性样品23份,阳性率约为32.0%(23/72),recomLine检出阳性样品24份,阳性率约为33.30%(24/72),检出阳性率两者差异不显著(P>0.05);在recomWell检出的23份阳性样品中,recomLine检测均为阳性,两者阳性检出符合率为100%(23/23);recomWell的漏检率约为4.1%(1/24)。结论 recomWellR和recomLine均可用于猪抗HEVIgG的检测,recomLine检测灵敏度略高于recomWell,且操作更简便,不需要特殊昂贵的检测设备,特别适合用于基层检测猪抗HEV IgG。     

Assessment of enzyme linked immunosorbent assay based diagnostic kits (Toxokit-G and Toxokit-M) for the detection of IgG and IgM antibodies to Toxoplasma gondii in human serum

An enzyme linked immunosorbent assay (ELISA) using penicillinase was developed in the form of diagnostic kits (Toxokit-G and Toxokit-M) for the detection of IgG and IgM antibodies to Toxoplasma gondii. The performance of both the kits was compared with commercially available diagnostic kits, i.e. Enzygnost-Toxoplasmosis/IgG (Behring Co., Germany), TOXOTEK-G (Flow Lab., U.K.) and Toxoplasma IgM Microassay (Diamedix Corp., U.S.A.) by testing toxoplasma-suspected human serum samples. The results indicate a good reliability between these diagnostic kits. Toxokit-G has 86.66 and 96.05% sensitivity and specificity respectively. The main advantage of Toxokit-G is that the end result can be assessed visually without using sophisticated instruments. Toxokit-M has 100% sensitivity and specificity and test results were not affected by the presence of antitoxoplasma IgG antibodies, rheumatoid factor or antinuclear antibodies.     

Evaluation of a cow-side immunoassay kit for assessing IgG concentration in colostrum

OBJECTIVE: To determine sensitivity and specificity of a cow-side immunoassay kit for assessing IgG concentration in colostrum. DESIGN: Prospective study. ANIMALS: 76 dairy and 11 beef cows of various parities. PROCEDURE: Colostrum from first, second, and third milkings and milk samples were collected, and IgG concentration was determined by means of radial immunodiffusion. The immunoassay was performed according to the manufacturer's instructions, and sensitivity and specificity were calculated by comparing results of the immunoassay (positive vs negative) with results of immunodiffusion (< 50 g/L vs > or = 50 g/L). RESULTS: 135 colostrum or milk samples were collected. Mean +/- SD colostral IgG concentrations, determined by means of radial immunodiffusion for dairy and beef cows were 65.4 +/- 51.4 g/L and 114.8 +/- 42.7 g/L, respectively. Mean IgG concentrations for first-, second-, and third-milking colostrum samples and for milk samples were 92 +/- 49.0 g/L, 74.6 +/- 45.1 g/L, 47.5 +/- 32 g/L, and 6.8 +/- 3.8 g/L, respectively. Sensitivity of the immunoassay (ie, percentage of samples with IgG concentration < 50 g/L with a positive immunoassay result) was 93%, and specificity (ie, percentage of samples with IgG concentration > or = 50 g/L with a negative immunoassay result) was 76%. CONCLUSIONS AND CLINICAL RELEVANCE: Results suggested that the immunoassay kit was an acceptable cow-side test to identify colostrum samples with IgG concentrations < 50 g/L. The immunoassay kit should be useful in screening colostrum for adequate IgG concentration before feeding to calves or storage.     

Quantification of bovine IgG, IgM and IgA antibodies to Clostridium perfringens B-toxin by enzyme immunoassay I. Preparturient immunization for enhancement of passive transfer of immunity

A quantitative competitive binding "triple-sandwich" enzyme immunoassay was developed and used to evaluated pathogen/class-specific antibody responses in Holstein-Friesian cows vaccinated against Clostridium perfringens B-toxin. Vaccination of cows at six weeks and again at two weeks prepartum increased pathogen-specific IgG levels in each dam's colostrum and respective calf's serum. Pathogen-specific IgG and IgM concentrations in dams' sera and colostra were related to subsequent pathogen-specific IgG and IgM neonatal sera concentrations. Only pathogen-specific IgA in dams' colostra was correlated to neonatal levels, possibly owing to a different origin and role of this immunoglobulin class. All class-specific colostral immunoglobulin levels were related to subsequent neonatal concentrations. Isotypic antibody responses against C. perfringens B-toxin were found with pathogen-specific IgM predominant in dams' sera and pathogen-specific IgA predominant in colostra and neonatal sera.     

Development of a disperse dye immunochromatographic test for the detection of antibodies against infectious bursal disease virus

For investigating the feasibility of using disperse dyes as an immunoassay chromogenic marker, a disperse dye, DADISPERSE NAVY BLUE SP, was selected in analyzing antibody against infectious bursal disease virus (anti-IBDV). With the color intensity revealed in the disperse dye immunochromatographic test (DICT) strip as the objective function, the optimal dyeing conditions were found as follows: dye concentration absorbance (at lambda(max)=587nm)=3, pH 7, 50 degrees C, for 10min. Under these conditions, the resultant dyed-antibody (rabbit anti-chicken) can produce an optimal color intensity reading of 55,054 on the strip. For performing qualitative immunoassay, chicken sera samples taken from different farms were used for the anti-IBDV titre assessment. The results of DICT strips showed very high sensitivity and specificity as compared to that analyzed by FlockChek enzyme linked immunosorbent assay (F-ELISA) kits. For quantitative immunoassay, it was found that the color intensity measured with DICT was linearly correlated to that of F-ELISA titre (r(2)=0.9687). Therefore, DICT was further applied to the detection of chicken anti-IBDV sera under vaccination in the farms. The average titres of the sampling groups exhibited a strong agreement to that of F-ELISA. Accordingly, the DICT method developed in this study, shown to be reliable, cheap and simple in both qualitative and quantitative immunoassays, is particularly suitable for point-of-need testing (PONT) in agricultural applications.     

人血清中分离纯化鸡IgG、IgM的实用方法

本试验用绵羊红细胞免疫鸡,免疫后5天采血,制备富集IgM血清。应用硫酸铵粗提、Sepharose-4B分子筛层析方法,从富集IgM的鸡血清一次性分离提取高纯度的IgM及纯度较高的IgG,DEAE－52纤维素离子交换层析进一步纯化,获得电泳纯的IgG。经鉴定该方法制备的IgG,IgM完全可以满足免疫学实验要求,从一份血清样品中同时纯化IgG、IgM两种免疫球蛋白,既省时又节省原料。     

猪血浆蛋白粉中IgG含量的酶联免疫检测方法研究

猪血浆蛋白粉已经成为断奶仔猪理想和必需的蛋白饲料来源。评价血浆蛋白粉质量的重要指标就是其中IgG的含量。研究通过制备猪IgG单克隆抗体,调试优化后建立了相应的酶联免疫检测方法。该方法最低检测浓度为10μg/g,阴性血浆蛋白粉添加10(1%)、100(10%)、200mg/g(20%)猪IgG标准品后用本方法测定回收率为73.1%～104.6%,批内、批间变异系数小于15%,可基本满足血浆蛋白粉质量监控需要。     

中草药复方添加剂对土杂鸡IgG的影响

将360只28日龄土杂鸡随机分为8组,在其日粮中分别按0．5％、1％、2％三个浓度添加自制中草药添加剂A和B,同时设抗生素组和对照组,于42、56、70日龄分别测定IgG 含量。结果显示:中草药添加剂能显著提高土杂鸡的IgG水平,且中草药添加剂A要优于B。