不同品种牛血清对3T3L1前脂肪细胞增殖与分化的影响

研究不同品种牛血清对3T3-L1细胞增殖与分化的影响,采用模型细胞体外培养法模拟牛脂肪组织生长环境,为牛大理石纹肉早期选择提供一种可能性的方法。抽取17头秦川牛和28头秦杂牛血清,先制备没灭活和灭活血清组培养细胞,MTT法检测细胞增殖相对数,油红O检测脂肪含量,再用不同品种牛血清培养细胞,检测细胞增殖相对数和脂肪含量,以及用比色法测定三磷酸甘油脱氢酶(GPDH)和脂肪酸合成酶(FAS)活性。结果表明,灭活血清培养细胞的增殖相对数量在分化培养第2和4天与分化脂肪含量在第2、4、6和8天都极显著高于没灭活血清组 (P＜0.01);秦杂牛血清培养细胞的数量在第2和4天显著高于秦川牛 (P＜0.05),秦川牛血清培养细胞分化的脂肪含量在第8天显著高于秦杂牛 (P＜0.05),其他天数没有显著性差异 (P＞0.05);分化第8天的细胞内GPDH和FAS酶活两牛种间没有显著性差异 (P＞0.05)。结果显示自制血清灭活比没灭活的更利于细胞的培养;牛血清品种是影响前脂肪细胞增殖与分化的因素,秦杂牛血清可能更有利于前脂肪细胞的增殖,而秦川牛血清可能更有利于前脂肪细胞的分化,但仍需进一步研究。     

间接酶联免疫吸附试验检测猪生殖与呼吸系统综合征血清抗体的研究

本研究建立了检测猪生殖与呼吸系统综合征（ＰＲＲＳ）血清抗体的间接酶联免疫吸附试验（ＥＬＩＳＡ），其抗原包被浓度为２μｇ／ｍｌ，血清最适稀释度为１∶１００。试验结果表明：ＥＬＩＳＡ的敏感性高于间接免疫荧光抗体试验（ＩＦＡ），是一种切实可行的诊断方法。     

羊群绵羊肺炎支原体抗体间接ELISA 检测方法的建立

利用绵羊肺炎支原体国际标准株Y-98,制备全细胞抗原,作为包被抗原,建立了一种间接ELISA方法。对4个不同羊场进行了血清学调查,确定了MoP抗体阳性率各为90%、70%、83.3%和73.9%。同间接血凝试验和鼻腔拭子培养结果相比,间接ELISA方法更加特异敏感,检出率更高,从而对该病的诊断和免疫程序的制定具有重要意义。     

Procedure for PEG treatment of bovine sera used for cell culture in virus diagnosis

A method for PEG treatment of bovine sera to be used for cell cultures is presented. The procedure consists in repeated freezing and thawing cycles of the sera, after PEG precipitation until a clear solution is obtained. The sera, so prepared, are homogeneous in composition and appropriate for cell growth in primary cell cultures and cell lines.     

抗伤寒杆菌Fe—SOD血清对志伤寒杆菌攻击小鼠的免疫保护作用

目的：制备兔抗伤寒杆菌Ｆｅ－ＳＯＤ血清,并研究其对鼠伤寒杆菌攻击小鼠的免疫保护作用,探讨ＳＯＤ与沙门氏菌致病性的关系。方法：用纯化的伤寒杆菌Ｆｅ－ＳＯＤ免疫家兔,制备兔抗ＳＯＤ血清,并用ＥＬＩＳ法检测抗血清效价;免疫电泳、双向琼脂扩散试验和抗血清对酶活笥抑制试验检测抗血清的特异性。将所制备的抗血清与鼠伤寒杆菌混合,放３７℃３０ｍｉｎ,经腹腔注入小鼠体内,并观察３天和７天小鼠的存活情况,以确定抗血清     

An enzyme-linked immunosorbent assay for detection of avian influenza virus subtypes H5 and H7 antibodies

Background

Avian influenza virus (AIV) subtypes H5 and H7 attracts particular attention because of the risk of their potential pathogenicity in poultry. The haemagglutination inhibition (HI) test is widely used as subtype specific test for serological diagnostics despite the laborious nature of this method. However, enzyme-linked immunosorbent assays (ELISAs) are being explored as an alternative test method.H5 and H7 specific monoclonal antibodies were experimentally raised and used in the development of inhibition ELISAs for detection of serological response specifically directed against AIV subtypes H5 and H7. The ELISAs were evaluated with polyclonal chicken anti-AIV antibodies against AIV subtypes: H1N2, H5N2, H5N7, H7N1, H7N7, H9N9, H10N4 and H16N3.

Results

Both the H5 and H7 ELISA proved to have a high sensitivity and specificity and the ELISAs detected H5 and H7 antibodies earlier during experimental infection than the HI test did. The reproducibility of the ELISA’s performed at different times was high with Pearson correlation coefficients of 0.96-0.98.

Conclusions

The ELISAs are a potential alternative to the HI test for screening of large amounts of avian sera, although only experimental sera were tested in this study.     

Mycoplasmas: use of polyvalent antisera for identification by indirect immunofluorescence.

It would be an advantage, under many circumstances, to be able to make use of polyvalent antisera in the process of identifying mycoplasmas. As the indirect immunofluorescence test is sufficiently sensitive and also generally accepted as being rather specific, this technique was chosen to investigate whether polyvalent antisera are applicable in routine identification of mycoplasmas. Three polyvalent sera were used, each consisting of 9 or 10 rabbit antisera raised against 29 of the more common species of the genus Mycoplasma. Twenty-six field strains were examined. One strain did not react with any of the 3 polyvalent antisera although it was later identified as M. bovigenitalium. The remaining 25 strains reacted with 1 and only 1 of the polyvalent antisera and were subsequently identified by immunofluorescence utilizing monospecific antisera. Strains of the following species were identified: M. arginini, M. bovigenitalium, M. bovis, M. bovoculi, M. canis, M. capricolum, M. cynos, M. edwardii, M. hominis, M. hyorhinis, M. molare, M. mycoides subsp. mycoides and M. opalescens. It is concluded that polyvalent antisera may be used in identification procedures and thereby permit the use of a limited number of monospecific antisera without preceding biochemical testing.     

衣原体单克隆抗体在羊血清诊断上的应用

采用淋巴细胞杂交瘤技术，研制出的抗衣原体单克隆抗体，应用酶联免疫吸附试验(ELISA)方法，对本地区所采用的698份羊血清进行检测，羊血清阳性检出率为22．06％，选择50份羊血清用已建立的检测衣原体抗体ELISA方法与传统的间接血凝试验(IHA)进行比较，其敏感性高。羊血清ELISA阳性检出率为26％，IHA为22％，前者比IHA高出4个百分点。     

BALB/c小鼠Doppel蛋白的原核表达及其抗血清的制备

利用已克隆的BALB/c小鼠Doppel蛋白基因,将其成熟蛋白编码区亚克隆至表达载体pGEX-6P-1上,构建重组表达质粒pGEX-6P-1-PRND。将重组蛋白表达载体转入E.coli BL21（DE3）中诱导表达,SDS-PAGE及Western blot分析结果表明,BALB/c小鼠Doppel蛋白在大肠杆菌中以包涵体形式高效表达。对表达的蛋白进行提取和纯化并免疫新西兰大白兔制备兔抗血清,初步建立了小鼠睾丸Doppel蛋白组织学分布的免疫组织化学方法。为进一步研究Doppel的结构、功能及其在TSEs发生发展中的作用提供基础材料和数据。