MD病毒抗独特型抗体的研制

制备鸡马立克氏病病毒(MDV)抗独特型抗体(Ab2),探讨Ab2模拟抗原诱导免疫应答的能力.用MDV免疫SPF鸡,分离提纯鸡抗MD-IgG(Ab1),Ab1免疫BALB/C鼠,用鼠脾细胞与SP2/0细胞制备Ab2杂交瘤细胞株,间接ELISA筛选阳性杂交瘤细胞株,经克隆、纯化2×株抗MDV独特型抗体杂交瘤细胞株(6E5,7A12),其中6E5培养上清、腹水ELISA效价分别为4000×、10000×,免疫20 d攻毒,具有抗MDV感染保护.     

非洲猪瘟病原学及单克隆抗体制备技术研究进展

非洲猪瘟是由非洲猪瘟病毒引起猪的高度接触性、传染性、出血性以及高死亡率的传染病。20世纪中期以来,已在非洲、欧洲和美洲等数十个国家流行,并在近几年内蔓延至欧亚两洲接壤处的格鲁吉亚、亚美尼亚、阿塞拜疆以及俄罗斯境内,其一旦侵入我国,将会给我国养猪业带来极大的危害。非洲猪瘟病原学研究以及制备相应的单克隆抗体对非洲猪瘟病毒快速诊断技术研究和疫苗研制有着重大的现实意义。主要从病原学和单克隆抗体制备方面对非洲猪瘟的研究进展进行了综述。     

马流感病毒双抗体夹心ELISA检测方法的建立

为建立一种快速、有效的检测马流感病毒(Equine influenza virus,EIV)的方法,以EIV中国分离株A/马/新疆/07(H3N8)制备的多克隆抗体为捕获抗体,原核表达的核蛋白(NP)制备的单克隆抗体为检测抗体,在国内首次建立了检测EIV的双抗体夹心ELISA方法.用该检测方法分别检测EIV、马动脉炎病毒、马疱疹病毒1型、马疱疹病毒4型和马乙型脑炎病毒阳性样品.结果表明,该ELISA方法具有良好的特异性;与常规检测EIV的血凝试验相比,其敏感性是后者的2.5～10倍;同时与H7N7亚型EIV有交叉反应.攻毒试验结果表明该方法可有效检测鼻腔分泌物中的EIV.该方法的建立为EIV的检测及早期防控提供了有效工具.     

间接ELISA检测猪伪狂犬病血清抗体

用猪肾传代细胞IBRS－2增殖猪伪狂犬病病毒（PRV）鄂A株，病毒培养上清液经硫酸铵沉淀、聚乙二醇（Mr20000）浓缩后作为包被抗原。用纯化的猪血清IgG免疫家兔，HRP村记撮的兔抗猪IgG，制备出高效价的酶标抗体，酶标抗体工作浓度为1：50000；经各种条件的选择，建立了检测猪伪狂犬病血清抗体的间接ELISA。所建立的间接ELISA抗原包被浓度为39.2mg/L，血清最佳稀释度为1：20，与猪细小病毒、猪、O型口蹄疫、猪衣原体标准阳性血清呈阴性反应，与标准阴性血清和临床未感染PRV的猪血清呈阴性反应；与猪伪狂犬病标准阳性血清、免疫猪血清和临床发病猪血清呈明显的阳性反应；与美国进口的PRV抗体检测ELISA诊断试剂盒检测结果比较，45份猪血清的阴、阳性检出符合率均为100％。表明建立的间接ELISA具有敏感性高、特异性强、重复性好的优点，可用于猪伪狂犬病血清抗体的定性和定量检测。     

Tracking fungi in soil with monoclonal antibodies

Species of the genus Trichoderma are ubiquitous soil-borne fungi that exhibit antagonism towards a number of economically important plant-pathogenic fungi and oomycetes. This review discusses recent developments in the use of monoclonal antibodies to detect these fungi in their natural soil environments and to quantify their population dynamics during antagonistic interactions with saprotrophic competitors in soil-based systems. Immunological approaches to detection and quantification are examined in relation to conventional plate enrichment techniques and to nucleic acid-based procedures. An example of recent research using a mAb-based assay to quantify the effects of saprotrophic competition on the growth of Trichoderma isolates in mixed species, soil-based, microcosms is presented. Future technological developments in immunoassays for tracking Trichoderma populations in soil are discussed and results presented showing the accurate detection and visualization of a plant growth-promoting isolate of T. hamatum in the rhizosphere of lettuce using mAb-based immunodiagnostic assays.     

Development and characterization of mouse monoclonal antibodies to eight human complement components: Analysis of reactivity with orthologs of nine mammalian genera

To study complement function in mammalian leishmanioses, we developed mouse monoclonal antibodies to the human complement system components C1q, C4, factor D, factor H, factor B, properdin, C5 and C9. Antibody specificity was determined by indirect and capture ELISA and by Western blot. In flow cytometry analysis, seven antibodies recognized the cognate component on human serum-opsonized Leishmania promastigotes. Antibody reactivity was screened against promastigotes opsonized with sera of nine mammalian genera: pig, guinea pig, goat, rabbit, cat, dog, hamster, jird and rat. No antibody recognized jird epitopes on promastigotes. Anti-C4, -properdin, and -C5b reacted with the orthologous protein of all other mammals tested except cat (anti-properdin) and hamster (anti-C5b); anti-C9 only recognized the rabbit ortholog, and anti-C1q, -factor B and -factor H did not react with any of the nine orthologs. Such interspecies crossreactive antibodies can be valuable tools for analysis of mammalian complement function in infectious diseases.     

产蛋鸡免疫程序对卵黄抗体效价的影响

试验用小鹅瘟免疫原按8种免疫程序接种120日龄产蛋鸡,分别在接种后不同时间琼脂扩散试验试验检测卵黄抗体效价,筛选出最佳免疫接种程序。基础免疫,产蛋鸡每只肌肉注射小鹅瘟活疫苗1.0 m L（5羽份）;在基础免疫后14 d进行第2次接种,加强免疫,产蛋鸡每只肌肉多点注射小鹅瘟灭活疫苗2.0 m L;在加强免疫后14 d进行第3次接种,强化免疫,产蛋鸡每只肌肉多点注射小鹅瘟灭活疫苗3.0 m L。     

Neutralising antibodies to wildebeest-derived malignant catarrhal fever virus in African wildlife

A total of 2,722 sera collected between 1963 and 1983, from 43 different species of wildlife in 11 African countries was examined for neutralising antibodies against the wildebeest-derived strain of malignant catarrhal fever (MCF) virus. Antibodies were demonstrated in 10 species of Bovidae which included eight species from the sub-family Hippotraginae and one species each from Bovinae and Antilopinae. Neutralising antibodies were also recorded in hippopotamus. It is suggested that the high prevalence of antibodies recorded in sera from waterbuck and reedbuck indicate infection with MCF. However, titres in other species may be due to antigenically related viruses.     

Effect of Repeated Use of Progestagen-PMSG Treatment for Estrus Control in Dairy Goats out of Breeding Season

Contents In order to analyze the effects of repeated use of progestagen-PMSG treatment, estrus and pregnancy results have been analyzed for 1989 in a Saânen dairy goat herd in which breeding takes place each year out of season after FGA/PMSG treatment. After the first 1989 treatment (169 goats), percentage of goats showing estrus and kidding have been lower for 59 multiparous than for 46 primiparous and 64 nulliparous females. Moreover, when 38 goats are treated for a second time in 1989, 44.7% exhibited estrus vs 71.0% after the first treatment (P < 0.05). The PMSG binding level before the 1st 1989 treatment is higher for multiparous (17.5 ± 23.1%) than nulli and primiparous (-0.06 ± 0.7 and 1.2 ± 1.9%) and is increased for all parities after treatment (23.2 ± 26.4 after vs 5.7 ± 15.0% before, P < 0.01). For nulliparous and primiparous females; PMSG binding levels are not different for pregnant or not pregnant nulliparous and primiparous goats. On the opposite, PMSG binding rates are higher in non pregnant (25.7 ± 23.3) than in pregnant multiparous goats (6.5 ± 15.9) (P < 0.01). However, when the binding rate is ≤ 5.12% (computerized distributions) multiparous goats exhibit estrus and pregnancy at levels not different from nulliparous or primiparous females (% estrus 95.8 vs 100 or 97.8%, % pregnancy 66.7 vs 70.3 and 63.0% respectively). Repeated use of PMSG during the female life or during one given year leads to active immunization against PMSG (as measured by percentage of binding of PMSG in plasma) decreasing the efficiency of ovarian stimulation out of breeding season. Inhalt: Einfluß einer wiederholten Progestagen/PMSG Behandlung zur Östruskontrolle außerhalb der Paarungssaison bei Milchziegen Um den Einfluß wiederholter Progestagen-PMSG-Behandlung zu prüfen, wurden Östrus und Trächtigkeitsergebnisse in einer Saanen Ziegenherde für das Jahr 1989 analysiert. Bei den untersuchten Tier en wird die Bedeckung jedes Jahr außerhalb der Saison nach FGA/PMSG-Behandlung vorgenommen. Nach der ersten Behandlung 1989 (169 Ziegen) war der Prozentsatz Ziegen, die einen Östrus zeigten und ablammten, bei 59 multiparen niedriger als bei 46 primiparen und 64 nulliparen Tieren. Darüber hinaus zeigten von 38 Ziegen nach einer zweiten Behandlung 1989 zu 44,7% einen Östrus gegenüber 71% nach der ersten Behandlung (p < 0,05). Die Bindungsfähigkeit für PMSG war bei der ersten Behandlung bei den multiparen Tieren höher (17,5 ± 23,1%) als bei nulli- und primiparen Tieren (-0,06 ± 0,7% und 1,2 ± 1,9%) und stieg für alle Tiere nach der Behandlung an (23,2 ± 26,4% nach zu 5,7 ± 15,0% vor der Behandlung, p < 0,01). Die PMSG Bindungsfähigkeit war nicht unterschiedlich bei tragenden und nichttragenden nulli- und primiparen Tieren. Dagegen waren die PMSG Bindungsraten bei nichttragenden Ziegen (25,7 ± 23,3%) höher als bei tragenden multiparen Tieren (6,5 ± 15,9%) (p < 0,01). Wenn allerdings die Bindungsraten unter 5,12% (Computerverteilung) sinkt, zeigen multipare Ziegen Östrus und Trächtigkeitsraten, die sich von multiparen und primiparen Tieren nicht unterscheiden (Östrus < 95,8% gegen 100 bzw. 97,8%; Trächtigkeit: 66,7% gegen 70,3 bzw. 63%). Wiederholter Gebrauch von PMSG, auch schon eine Applikation innerhalb eines Jahres führt zur Immunisierung gegenüber PMSG (gemessen an dem Prozentsatz Bindungsvermögen im Plasma) und senkt die Effektivität der ovariellen Stimulation außerhalb der Saison.     

抗猪支原体共同抗原单克隆抗体的制备与鉴定

以猪肺炎支原体（Mycoplasma hyopneumoniae,Mhp)168菌株F332作为抗原，免疫8周龄BALB/c小鼠，利用淋巴细胞杂交瘤技术，获得两株能稳定分泌特异单克隆抗体的杂交瘤细胞株。两株单抗与Mhp和猪鼻支原体（M.hyorhinis,Mh)等反应，而不与鸡支原体，对照血清等反应。结果表明两株单抗可能是针对猪支原体共同抗原的单抗，用SDS-PAGE电泳和Western-blot分析猪支原体的膜蛋白成分。结果显示，猪支原体的共同抗原是80kd和30kd蛋白，两株单抗均与Mhp和Mh的30kd蛋白条带反应，表明这两株单抗特异地针对猪支原体的共同抗原成分30kd蛋白，可以看出，这两株单抗在Mhp的抗原分析，血清学诊断和疫苗质量监测以及猪源支原体污染细胞检测等方面有重要应用价值。