Differential reactivity of human serums with early antigens induced by Epstein-Barr virus

Inoculation of 64-10 or Raji cultures with Epstein-Barr virus derived from the HRI-K clone of the P3J Burkitt's lymphoma line caused abortive infections in most of the lymphoblastoid cells with synthesis of "early antigens" but few, if any, capsids. Antibodies to early antigens were detected by indirect immunofluorescence in serums of many patients with infectious mononucleosis, Burkitt's lymphoma, or nasopharyngeal carcinoma. These antibodies were rarely present in other serums even though some of them showed high titers of antibodies to Epstein-Barr virus when assayed on EB3 Burkitt tumor cells; they also prevented synthesis of early antigens, provided the serums were mixed with the virus prior to inoculation. Antibodies to early antigens possibly reflect current or recent disease processes that are associated with the virus.     

Hepatitis B virus antigen: validation and immunologic characterization of low-titer serums with 125 I-antibody

Radioimmunnoassay was used to determine the serologic subspecificities of 85 blood donor serums positive for hepatitis B virus-associated antigen. There was complete agreement with results obtained by immunoprecipitatiotn of 43 serums. The remaining 42 serums were negative by immunoprecipitation but followed type-specific immunology by radioimmunoassay, and this served as a validation for authentic hepatitis B virus antigen.     

Hemagglutination assay for antigen and antibody associated with viral hepatitis

Hemagglutination assays are described for measuring hepatitis-associated Australia antigen and antibody. Red cells coated with isolated antigen, with chromic chloride as a coupling agent, are used for detection of antibodies. Detection of the antigen in serums depends on inhibition of hemagglutination. The test has the sensitivity and rapidity of the best tests available, is simpler to perform, and lends itself to large-scale screening of blood donors.     

鸡新城疫-传染性支气管炎-H9亚型禽流感三联灭活疫苗的制备及检验

[目的]制备鸡新城疫-传染性支气管炎-H9亚型禽流感[ND-IB-AI（H9亚型）]三联灭活疫苗。[方法]以NDV La Sota株、IBVM41株、AIV（H9亚型）WD株为毒种,分别接种鸡胚,制备NDV、IBV及AIV（H9亚型）抗原液;采用病毒超滤系统浓缩病毒抗原液,并用甲醛溶液进行灭活;将浓缩灭活后的3种抗原液按一定比例混合（1∶1∶1）,以司本80及吐温80为乳化剂,10号白油为佐剂,乳化制成ND-IB-AI（H9亚型）三联灭活疫苗;对制备的疫苗进行无菌检验和物理性状观察。[结果]共制备了3批ND-IB-AI（H9亚型）三联灭活疫苗实验室制品,经无菌检验为阴性,外观为乳白色,剂型为油包水型（W/O型）,粘度6.36.8 s,离心和37℃放置21 d均不分层。[结论]所制备3批ND-IB-AI（H9亚型）三联灭活疫苗均无细菌污染,其物理性状均能达标。     

口蹄疫病毒3AB试剂盒的检测及比较

以重组口蹄疫病毒非结构蛋白3AB为检测抗原,研制了口蹄疫病毒非结构蛋白ELISA检测试剂盒,该试剂盒可用于判定被检动物是否感染口蹄疫病毒。重组口蹄疫病毒非结构蛋白3AB作为检测抗原包被酶标板反应孔,以辣根过氧化物酶标记的重组蛋白A/G作为第二抗体,建立了抗非结构蛋白抗体检测的方法。结果表明,在1 746份免疫猪血清中,3AB-ELISA对免疫猪血清的特异性为98.68%;在1 077份健康非免疫猪血清中,3AB-ELISA对健康非免疫猪血清的特异性为98.68%;在36份人工感染猪血清中,3AB-ELISA对阳性检出率为100%。而牛的特异性的各项指标和阳性检出率分别为94.04%,97.96%和100%。猪/牛口蹄疫病毒非结构蛋白酶联免疫吸附试验检测试剂盒与国外同类试剂盒比较,阳性检出率高出12.5个百分点。     

Interactions of immunoglobulins G and M in the detection of the mammalian C-type virus cross-reactive antigen

The mammalian C-type tumor viruses share an antigenic determinant, gs-3, located on the major internal polypeptide of the virion. Detection of this determined in gel diffusion assays by antiserums prepared in rats by immunization with rat tumor homogenates carrying murine virus and serums prepared in a rabbit by immunization with purified murine gs antigen depended on antibodies present in the fractions containing immunoglobulins M and G. The immunoglobulin G fraction by itself precipitated only the homologous murine antigen. Neither fraction alone precipitated heterologous (cat, rat, or hamster) antigen (definition of the gs-3 reaction), while a mixture of the two fractions did. The gs-3 reaction was eliminated by treatment of the serums with beta-mercaptoethanol, also indicating a requirement for immunoglobulin M antibodies.     

利用Asia1型口蹄疫病毒VP1蛋白的单克隆抗体建立单抗竞争ELISA方法

1材料与方法1．1材料新生牛血清购自GIBICO生物公司;青霉素、链霉素购自哈尔滨制药六厂;降脂烷购自sigma公司;过碘酸钠购自上海生物试剂公司;脱脂乳购自飞鹤乳业公司;Babl／c小鼠购自兰州生物制品研究所。     

Hemadsorption-negative plaque test: new assay for rubella virus revealing a unique interference

A simple and rapid plaque procedure has been developed for detecting and accurately assaying rubella virus in a noncytopathic virus-cell relationship. Plaque-formation is based on the development, in individual cells infected with rubella virus, of a unique type of intrinsic interference to infection with Newcastle disease virus. Rubella virus-infected cells challenged with Newcastle disease virus and tested for hemadsorption 15 hours later stand out as hemadsorption-negative areas. Individual living cells infected with rubella virus can be resolved under conditions allowing standard cloning procedures. In principle, the hemadsorption-negative plaque test can be used to search for a new class of noncytopathic, non-hemadsorbing viruses-those that induce an intrinsic interference to infection by any hemadsorbing virus.     

利用Asia 1型口蹄疫病毒VP1蛋白的单克隆抗体建立单抗竞争ELISA方法(英文)

Using the purified VP1 protein of Asia 1 type foot-and-mouth disease virus as the antigen, the purified monoclonal antibody was labeled by the sodium periodate method and the monoclonal antibody competitive ELISA was established in this study. Ten positive porcine foot-and-mouth disease serums and more than two hundreds negative serum were tested, and the results were the same as the background of samples. The sensitivity test and replicate test indicated that this method was stable and sensitive, which was suitable for monitoring Asia 1 type porcine foot-and-mouth disease virus antibody.     

新城疫病毒纯化方法的比较研究

为了比较研究红细胞吸附释放法、PEG-6000沉淀法和超速离心法对新城疫病毒的纯化效果,将含有NDV的鸡胚尿囊液分别用上述3种方法处理,并测定血凝(HA)效价。结果表明,红细胞吸附释放法中采用50%的醛化红细胞纯化病毒效果最佳;PEG-6000沉淀法中,用9%的PEG-6000加1%NaCl处理病毒后再沉淀2~4 h效果最佳;采用超速离心法纯化的病毒主要分布在40%和50%蔗糖区带之间。3种方法比较,以超速离心法效果最好,红细胞吸附释放法和PEG-6000沉淀法纯化效果差异不大。     

鸡抗病促生长剂的试验研究

采用ＤＮＡ重组技术和蚀斑技术获得融合基因的重组病毒，制备了鸡生长抑制因子（ＳＳ）免疫原，与鸡传染性ＩＢＤ、ＮＤ免疫原共同高免健康产蛋鸡，使其卵黄中产生高效价的抗鸡ＳＳ、ＩＢＤ、ＮＤ抗体，利用含高效价的卵黄制成鸡抗病促生长剂。实验结果表明，该制剂对人工感染的ＩＢＤ鸡治愈率为９７．５％，预防保护率９８．４％；对流行区自然感染的ＩＢＤ鸡治愈率９６％，预防保护率９７％。对健康肉仔鸡应用，可使其育成出栏时间提前３天，每只平均比对照增重１３５．７ｇ；饲料转化率提高８．３７％。     

新城疫病毒HN蛋白结构及其生物学活性研究进展

新城疫病毒(Newcastle disease virus,NDV)HN糖蛋白位于病毒囊膜表面,与病毒的毒力及致病性密切相关,因此在NDV的研究中具有重要的意义。从HN蛋白的1级结构、空间结构分析了与其相应的受体结合能力、神经氨酸酶活性和促膜融合活性等生物学活性。     

新城疫病毒HN蛋白的抑瘤机制及其研究进展

新城疫病毒囊膜上较大的纤突样糖蛋白由新城疫病毒血凝素-神经氨酸酶(HN)基因编码。HN蛋白即血凝素-神经氨酸酶蛋白,它不仅介导病毒吸附于肿瘤细胞,而且与融合蛋白一起帮助病毒侵入肿瘤细胞,其神经氨酸酶活性能水解宿主细胞表面的唾液酸,暴露宿主细胞的生物识别位点。它还能够水解病毒粒子表面糖链末端的唾液酸,防止病毒粒子在宿主表面簇集,并且能够引入新的粘附因子和免疫原性分子,诱导机体免疫系统识别。     

Serum hepatitis antigen (SH): rapid detection by high voltage immunoelectroosmophoresis

An immunoelectroosmophoretic technique for rapid detection of the antigen (SH) associated with the serum hepatitis virus has been devised. The technique maintains the specificity characteristic of the Ouchterlony gel-diffusion method, yet detects in 1 to 2 hours one-tenth the amount of antigen required for gel diffusion. The test has immediate application to blood-banking practice since it permits the screening of such labile products as platelets and fresh whole blood, and the detection of antigen in additional serums negative by the Ouchterlony technique.     

新城疫病毒毒力的演化研究进展

新城疫是由新城疫病毒引起禽的一种急性、热性、败血性和高度接触性传染病.该病主要以高热、呼吸困难、下痢、神经紊乱、黏膜和浆膜出血为特征.新城疫对我国养禽业的发展造成了巨大的威胁,其病毒是一种突变快、进化快的病原,该病毒毒力的强弱并不是一成不变的,其毒力具有本身的演化趋势.通过对新城疫病毒的生物特性、新城疫病毒毒力的演化及与其毒力演化相关的因素研究,旨在探讨新城疫病毒毒力的演化与流行趋势的关系.     

鸡新城疫病毒F基因部分片段克隆表达及其抗体制备

[目的]为进一步研究新城疫病毒的分子结构和基因疫苗奠定基础。[方法]用自行设计的l对引物,通过RT-PCR从接毒的SPF鸡胚的尿囊液中克隆了新城疫病毒F基因部分片段,将该基因片段插入含谷胱甘肽(GST)基因的质粒pGEX-4T-1,构建了重组质粒,对提取的融合蛋白进行亲和层析纯化和琼扩、ELISA及W estern-blot检测。[结果]通过克隆和PCR扩增获得新城疫病毒F基因的部分片段,大小为509 bp;对构建的重组质粒pGEX-4T-1-F509经诱导表达,获得分子大小约为44 000 bp的融合蛋白,其中F基因的部分片段大小约18 000 bp;经亲和层析,获得纯化GST-F509融合蛋白,进一步用该蛋白免疫小鼠,制备了鼠源抗鸡新城疫病毒F蛋白抗体。[结论]琼脂扩散试验、ELISA及W estern-blot检测表明,该融合蛋白中的F片段具有良好的抗原性。     

鹅源新城疫病毒M基因的原核表达及其多克隆抗体制备

试验旨在制备原核表达鹅源新城疫病毒(NDV)JS/5/05/Go株M蛋白的多克隆抗体并进行鉴定。以鹅源NDV总RNA为模板,RT-PCR扩增M基因,亚克隆到原核表达载体pET-32a(+)获得重组质粒pET32a-M,转化E.coli BL21(DE3)菌株进行诱导表达,SDS-PAGE电泳检测表达产物;用KCl染色切胶纯化法纯化重组蛋白;采用切胶免疫小鼠的方法制备M蛋白多克隆抗体,抗体效价用间接ELISA检测,特异性用Western blot和间接免疫荧光法鉴定。结果表明,在大肠杆菌中成功表达了分子量约为55 000的重组蛋白,用切胶纯化法获得了纯度较高的重组蛋白;ELISA法检测抗体效价可达1∶102 400,Western blot和间接免疫荧光试验结果表明制备的多克隆抗体能够特异性识别纯化的M蛋白及NDV自身表达的M蛋白。     

新城疫病毒F_(48)E_9株重组NP蛋白单克隆抗体制备及鉴定

制备新城疫病毒(NDV)NP蛋白单抗,以期为NDV抗原表位研究及检测方法建立方面奠定基础。利用NDV F48E9株pET32a-NP重组蛋白免疫BALB/c小鼠,采用杂交瘤细胞技术,间接ELISA筛选,获得了2株稳定分泌抗NP重组蛋白单克隆抗体的杂交瘤细胞株,分别命名为1G3、4B12。经间接ELISA测定,腹水抗体效价分别为1 2.56×105、1 5.12×105。亚类鉴定结果表明这两株单抗均为IgG1。Western blot分析结果显示,1G3、4B12均能特异性识别重组NP蛋白。与NDV感染细胞经间接免疫荧光试验检测均呈黄绿色荧光。经相加ELISA测定表明两株单抗识别的抗原表位不同。     

Radioimmunoassay for human procollagen

Rabbit antiserums were produced against the procollagen molecule secreted into the medium of cultured human skin fibroblasts. The isolated antigenic, amino terminal portion of the procollagen molecule was purified, labeled with iodine-125, and used in a radioimmunoassay which detected nanogram quantities of the same antigen. With the assay, immunologically identical molecules were detected in the culture mediumn of different strains of human fibroblasts and in normal human serums. Serumns from human cord blood contained a 12-fold higher concentration of the antigen than serums from adults, while serums other vertebrates gave reactions to incomplete cross-reactivity or non-reactivity.     

鸡新城疫的诊断及病理分析

[目的]对疑似鸡新城疫病例进行诊断,从而对防控该病提供依据。[方法]应用常规病理剖检技术解剖死亡鸡只,观察病理变化。病死鸡组织脏器研磨等处理后接种10日龄SPF鸡胚,取尿囊液进行平板血凝试验（HA）与平板凝集抑制试验（HI）。取病鸡肠道、腺胃、脑、十二指肠、肺脏等组织制作石蜡切片,HE染色,镜检。[结果]HA和HI结果显示,收集的尿囊液可凝集鸡红细胞,同时这种凝集作用又可被抗新城疫阳性血清抑制。病理剖检可见,各组织脏器有明显的出血性病变,实质细胞均出现不同程度的损伤。[结论]该分离毒株被确诊为鸡新城疫病毒。