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。     

Serum auto antibodies and clinical/pathological features in German shepherd dogs with a lupuslike syndrome

This study presents 8 dogs of German Shepherd breed (6 males, 2 females, 2-5 years of age at onset of the disease) with a lupus like syndrome characterized by febrile polyarthritis, wasting, nephropathy, cutaneous lesions and high positive titres of ANA (antinuclear antibodies) of speckled type. The serum autoantibodies were further characterized by double immunodiffusion against ENA (extractable nuclear antigen), ELISA for Histone antibodies (Histon fraction H-24A and H-3S), indirect IF on rat-liver sections, non treated and RNase/DNase digested sections for DNP/RNP antibodies, and smears of a hemoflagellate C. luciliae for antibodies vs doubbel strained DNA, (dsDNA). Thus, the high ANA titres in these dogs represent varying types of autoantibodies against nucleoproteins of both DNA and RNA nature, associated histone antigens and non-histone antibodies (RNA and Sm) as well. Rheumatoid Factor titres in serum from these dogs were low or negative. Immunoglobulin deposits at dermo-epidermal junctions were demonstrated in some of the dogs with hyperkeratotic skin lesions. High concentration of serum-IgG was a constant finding in combination with anemia and in most cases leukopenia probably related to the chronic inflammatory process in these animals. Autoimmune hemolytic anemia (AIHA) or thrombocytopenia was not detected in these dogs.     

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.     

对2005年玉米雌穗分化中几种异常现象的分析

2005年辽宁省丹东地区玉米生产在较大范围内发生了雌穗分化过程中的异常现象，涉及的品种之多，面积之大，在历史上也是罕见的。由于这种异常现象大面积发生，从而造成东港市玉米大幅度减产，减产幅度达30％～40％，个别地块达50％。本文将这些异常现象加以分析、整理，并对玉米雌穗分化过程中几种异常现象的表现、规律、成因和对策进行了初步探讨。     

斑点金标免疫渗滤法诊断家畜血吸虫病的敏感性和特异性

研制成家畜血吸虫病斑点金标免疫渗滤法（DIGFA）诊断试剂盒可测出人工感染血吸虫14—42d的病牛、猪和兔的血清及血纸抗体，比病原体检查法提早28d查出病畜，达到早期诊断的要求；同时亦能检出仅感染1对血吸虫病牛抗体，证实DIGFA具有较高的敏感性。血吸虫与肝片吸虫、锥虫、蛔虫、囊虫和旋毛虫病之间未见交叉反应现象，说明本法具有较强的特异性。     

抗猪瘟病毒单克隆抗体的研究——Ⅱ抗猪瘟兔化弱毒单克隆抗体杂交瘤细胞株的建立

用聚乙二醇沉淀浓缩纯化猪瘟兔化弱毒中国株(SFV-C)免疫 BALB/C 小鼠,取其脾脏,制备淋巴细胞,与 FO 骨髓瘤细胞融合,经 EliSA 检测和有限稀释法克隆化,最后筛选出5株对SFV 的单克隆抗体杂交瘤细胞.所制成的腹水抗体,一株只对 SFV-C 发生特异性反应,四株与SFV-S,SFV-F 及 BVDV oregon 发生微弱的交叉反应.细胞上清液的 EliSA 效价为1:800～1600,腹水效价为1:10~6～10~7.     

Production and Identification of High Affinity Monoclonal Antibodies Against Pesticide Carbofuran

To produce high-affinity monoclonal antibodies against pesticide carbofuran, and the develop immunochemical assays for people＇s health and environmental protection, the hapten 4-[[（2,3-dihydro-2,2-dimethyl-7-benzofuranyloxy） carbonyl]- amino]-butanoic acid （BFNB） of carbofuran was synthesized and Balb/c mice were immunized by the hapten-carrier （BFNB-bovine serum albumin, BFNB-BSA） conjugates. The splenocytes of immunized mice were fused with Sp2/0 cells and the cultural supernatants of hybridoma cells were screened by the indirect enzyme-linked immunoabsorbent assay （ELISA）, based on BFNB-ovoalbumin conjugates （BFNB-OVA）. Purified monoclonal antibody （McAb） was obtained from fluids of ascites, deposited by octanoic acid and ammonium sulfate. The affinity and the specificity of McAb were characterized by ELISA or indirect competitive ELISA. A hybridoma cell line （5D3） secreting anti-carbofuran McAb had been established. The titer of culture medium and ascites was up to 1：2.048 × 10^3 and 1：1.024 × 10^6, respectively, and the subtype of the McAb was IgG1. The affinity constant of the McAb was about 2.54 × 10×9 L mol^-1, with an IC50 value of 1.18 ng mL^-1 and a detection limit of 0.01 ng mL^-1. Cross-reactivity studies showed that the McAb was quiet specific for carbofuran, as among the four analogous compounds, they were all hardly recognized （4.59 × 10^-4% for 2,3-dihydro-2,2- dimethyl-7-benzofuranol and less than 3.0 × 10^4% for others）. The prepared McAb had a very high affinity and specificity, and it could be used to develop ELISA for rapid determination of carbofuran.     

抗沙门氏菌O_4、O_(12)抗原特异单克隆抗体的产生及其特性鉴定

用鼠伤寒沙门氏菌的热灭活全菌和热酚—水抽提脂多糖抗原免疫BALB/c小鼠,取脾脏淋巴细胞与SP_2/O细胞融合,获得稳定分泌抗沙门氏菌O_4和O_(12)抗原的特异单克隆抗体的杂交瘤细胞系7个,分别命名为AG24-1、AG90-5、BG1-8、BG17-12、BG66-10、BG78-15、EG19-11。小鼠腹水中的单抗直接凝集滴度为10~2-10~4,间接免疫荧光滴度为10~3-10~6。与沙门氏菌和肠杆菌科中其他细菌的反应性试验表明,BG78-15是O_(12)因子特异单抗,其他6种为对O_4因子的特异单抗。     

新城疫病毒感染绵羊诱导免疫应答反应的实验研究

为探索新城疫病毒(NDV)作为羊病毒病疫苗载体的可行性,验证NDV感染绵羊的可能性,本研究采用NDV Clone-30株分别通过滴鼻和气管灌注两种途径接种绵羊,接种后观察临床症状,检测接种动物排毒情况,通过血凝抑制试验检测接种绵羊特异性血凝抑制抗体,ELISA试验检测接种绵羊血清特异性IgG抗体,微量中和试验检测接种绵羊血清中和抗体.研究结果表明,NDV在绵羊体内是非致病性的,并且通过两种接种途径在绵羊体内均可以产生血凝抑制抗体、NDV特异性IgG抗体和中和抗体反应.本研究结果表明NDV有可能作为宿主限制性疫苗载体用于预防无特效疫苗的羊类疾病,且与气管灌注接种途径比,滴鼻接种途径相对安全.