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荧光抗体

荧光抗体的相关文献在1982年到2022年内共计177篇,主要集中在畜牧、动物医学、狩猎、蚕、蜂、内科学、预防医学、卫生学 等领域,其中期刊论文144篇、会议论文11篇、专利文献100543篇;相关期刊94种,包括微生物学杂志、中国麻风皮肤病杂志、实验动物与比较医学等; 相关会议9种,包括第十三次全国养犬学术研讨会、中国畜牧兽医学会兽医病理学分会第十六次、中国病理生理学会动物病理生理专业委员会第十五次学术研讨会、中国畜牧兽医学会动物传染病学分会第三届猪病防控学术研讨会等;荧光抗体的相关文献由533位作者贡献,包括文明、朱丽萍、颜世敢等。

荧光抗体—发文量

期刊论文>

论文:144 占比:0.14%

会议论文>

论文:11 占比:0.01%

专利文献>

论文:100543 占比:99.85%

总计:100698篇

荧光抗体—发文趋势图

荧光抗体

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  • 文明
  • 朱丽萍
  • 颜世敢
  • 周碧君
  • 夏咸柱
  • 张少鹏
  • 张贯京
  • 徐春志
  • 戴爱玲
  • 李慧玲
  • 期刊论文
  • 会议论文
  • 专利文献

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    • 左志丽
    • 摘要: 近几年,猪肉养殖产业的规模化、科学化发展,在保证了猪肉供应稳定性的同时,由于养殖密度的不断提升与不同猪种引进等方面的问题,使得猪传染病的预控难度不断提升。猪瘟作为影响养猪产业经济效益的传染疾病之一,该病的临床表现具有非典型、隐形感染等特点。基于此,本文主要通过对实验室猪瘟诊断的荧光抗体、猪瘟酶标记抗体等技术的分析,对猪瘟抗体监测对照实验与判定标准进行探析。
    • 左志丽
    • 摘要: 近几年,猪肉养殖产业的规模化、科学化发展,在保证了猪肉供应稳定性的同时,由于养殖密度的不断提升与不同猪种引进等方面的问题,使得猪传染病的预控难度不断提升.猪瘟作为影响养猪产业经济效益的传染疾病之一,该病的临床表现具有非典型、隐形感染等特点.基于此,本文主要通过对实验室猪瘟诊断的荧光抗体、猪瘟酶标记抗体等技术的分析,对猪瘟抗体监测对照实验与判定标准进行探析.
    • 中国动物疫病预防控制中心1
    • 摘要: 由于非洲猪瘟(ASF)缺乏预防用疫苗,为防止疫病的传播,就需要实施严格的卫生和生物安全控制措施,而这就依赖于疫病的快速、可靠的早期诊断。ASF的诊断是指确诊动物正在感染或者曾经感染过非洲猪瘟病毒(ASFV)。因此,适用的诊断技术包括检测和识别ASFV特异性抗原、DNA或抗体的技术,所获取的检测信息也是控制和根除计划的重要保障。在选择诊断技术时,分析疫病感染期非常重要(图1)。由于感染动物所处的感染期不同,因此在疫情和控制/根除计划中,需要同时检测病毒和抗体以确保准确性。
    • 于友水
    • 摘要: 本病发病急,病程短,临床诊断困难。调查该地区有关炭疽发生及尸体处理情况,有助于本病的诊断,但真正确诊必须依靠细菌学和血清学诊断。病死畜若有炭疽可疑,切不可剖检。可剪耳尖一小块,然后局部做适当处理(如烧烙),或用消毒棉棒蘸天然孔流出的血液少许,置灭菌试管中送检。1镜检镜检诊断炭疽有重要价值,但必须采取发病期及刚刚死亡动物的血液或其他材料制成涂片,用多色性美蓝、瑞氏染液或姬姆萨染液染色。
    • 乔绪稳; 张元鹏; 陈瑾; 张浩明; 侯立婷; 杨利; 郑其升; 侯继波
    • 摘要: Our experiment was to express recombi-nant antibody fused with EGFP fluorescent in E.coli for de-tecting porcine circovirus type 2( PCV2) in direct immu-nofluorescence assay ( DIF). The plasmid pET28a- V22-EGFP vector was transformed into Escherichia coli BL21 (DE3) and V22-EGFP protein was induced by IPTG. Ex-pression and solubility of recombinant V22-EGFP protein was analyzed by SDS-PAGE and Western blot and purified by Ni-NTA His?Bind Resin. The purified V22-EGFP protein was used to establish PCV2 direct immunofluorescence assay. SDS-PAGE and Western blot results revealed that recombi-nant V22-EGFP protein was successfully expressed with 60% soluble protein. With a obviously green appearances the puri-fied V22-EGFP protein showed great and stable fluorescence activity, even stored at -20 °C for 6 months. The DIF assay based on V22-EGFP showed good specifity to PCV2-infected PK-15 cells only. Compared to the conventional PCV2 IFA method, the PCV2 DIF based on recombinant V22-EGFP was more stable, sensitive and specific, so it enjoys a great poten-tial application in PCV2 research.%本研究旨在利用原核大肠杆菌系统表达1株猪圆环2型病毒(Porcine circovirus type 2,PCV2)小分子荧光抗体,以及该荧光抗体用于PCV2病毒直接免疫荧光(Direct immunofluorescence assay,DIF)检测.选取1株筛选到的PCV2特异性纳米抗体基因V22,按照E.coli原核表达系统进行密码子优化重新合成,与荧光蛋白基因融合后插入pET28(a)中,20°C条件下诱导表达16 h,利用SDS-PAGE、Western-blot技术鉴定蛋白是否表达及其可溶性,并对可溶荧光抗体蛋白进行镍柱亲和层析纯化,纯化蛋白用于建立PCV2特异性DIF检测方法,同时对其工作条件、工作浓度、特异性以及稳定性进行验证,并与PCV2经典间接免疫荧光检测方法进行比较,验证其灵敏度.结果表明,PCV2荧光抗体基因在E.coli原核表达系统中表达,可溶蛋白占目的蛋白的60%左右;纯化后荧光抗体蛋白呈现明显绿色,具有良好的荧光活性,不同批次表达及纯化的荧光抗体效果一致,-20°C保存6个月后抗体效价不变;PCV2特异性DIF试验结果表明该荧光抗体只对PCV2病毒感染细胞具有良好的反应原性,利用该抗体建立的直接免疫荧光法能够用于PCV2病毒滴度的测定,滴度测定结果与间接免疫荧光法检测结果一致.说明,在原核系统中成功表达PCV2的小分子荧光抗体,该荧光抗体蛋白具有良好的特异性、灵敏度及稳定性,用该抗体建立的PCV2特异性DIF操作简单,且具有较好的特异性和敏感性,对PCV2的检测有潜在应用价值.
    • 徐黎晖; 彭忠; 赵婷婷; 徐守兴; 陈焕春; 吴斌
    • 摘要: To develop a rapid,sensitive,and accurate direct-immunofluorescence method (DFM) detecting porcine pseudorabies virus (PRV),a fluorescein isothiocyanate-marked monoclonal antibody (FITC-MAb) against the gE protein of PRV was prepared and tested in this study.The results showed that the F/P ratio of the FITC-MAb was 2.578±0.012,which corresponds to the standard requirement.The best working conditions for the method were that the cells were immobilized with 60% acetone and 40% ethanol at-20 °C for 20 min,marked with 15 μg/mL FITC-MAb,and incubated at 37 °C for 1 h.Sensitivity tests showed that the method we established could detect PRV at 107-fold of dilution (1 ×TCID50).The FITC-MAb we prepared only displayed positive reaction with PRV wild type strains Ea and CH/SMX/2012,but had no cross reaction with PRV vaccine strain Bartha-K61,and other pathogenic viruses including classical swine fewer virus (CSFV),porcine parvovirus (PPV),porcine circovirus type 2 (PCV2) and porcine reproductive and respiratory syndrome virus (PRRSV),suggested the method had good specificity.Preliminary application of the method detecting 83 clinical samples showed that the accordance rate of DFM and PCR was 74.7%.Particularly,the accordance rate between the two methods detecting the samples of lymph node and tonsil was 85.7% and 93.3%,respectively.Our results indicated that the PRV direct immunofluorescence method established in this study layed the foundation for clinical PRV detection and complex diagnosis,which was suitable for clinical application.%为建立一种快速、敏感、准确的猪伪狂犬病病毒(PRV)直接免疫荧光检测方法(DFM),本研究通过制备并纯化PRV gE蛋白单克隆抗体(MAb),将其标记异硫氰酸荧光素(FITC)后对荧光抗体的最佳工作条件进行摸索;并对荧光抗体的特异性、敏感性、重复性和稳定性进行测定;应用DFM对从临床收集的83份疑似病料进行检测,同时用PCR检测作为对照,计算符合率.结果显示,本研究制备FITC标记抗体的F/P比值为2.578±0.012,符合标准;荧光抗体的最佳工作条件为:使用固定剂(60%丙酮+40%乙醇)于-20°C固定20 min,抗体工作浓度为15μg/mL,抗体孵育时间为1h;荧光抗体能在107病毒稀释度(1×TCID50)检出阳性信号,与PCV2、PPV、PRRSV、CSFV、PRV Bartha-K61均无交叉反应;批内和批间重复性良好,-20°C保存5个月染色后荧光强度仍稳定;83份临床样品检测结果显示,DFM与PCR的符合率为74.7%,其中淋巴结和扁桃体的符合率较高,分别为85.7%和93.3%.本研究建立的PRV直接免疫荧光检测方法为临床PRV检测和复合诊断奠定基础,适用于临床推广.
    • 何辉; 乔绪稳; 陈瑾; 于晓明; 张元鹏; 侯继波; 郑其升; 孙卫东
    • 摘要: [目的]本文旨在研究大肠杆菌表达的猪瘟病毒(CSFV)荧光抗体用于病毒直接免疫荧光检测的可行性。[方法]在基因水平上对编码猪瘟病毒纳米抗体(VHH)与绿色荧光蛋白(EGFP)的融合基因进行密码子优化,优化后的融合基因插入p ET32a(+)载体并在大肠杆菌BL21(DE3)中融合表达。表达的融合蛋白经过SDS-PAGE、Western blot进行表达及可溶性鉴定。利用镍柱亲和层析获得纯化的重组蛋白,经浓缩后用于猪瘟病毒直接免疫荧光检测并与CSFV间接免疫荧光试验进行效果比较,将该荧光抗体孵育CSFV、伪狂犬病病毒(PRV)、猪细小病毒(PPV)和猪圆环病毒2型(PCV-2)感染的PK15细胞从而鉴定荧光抗体的特异性。[结果]SDS-PAGE结果表明荧光抗体在大肠杆菌中实现部分可溶性表达。特异性试验结果表明,只有接种CSFV的PK15细胞组出现明显的绿色荧光,而其他组不出现或只出现少量非特异性荧光。与用商品化单抗建立的间接免疫荧光试验结果相比,该荧光抗体建立的直接免疫荧光方法具有更强的清晰度。[结论]猪瘟荧光抗体可在大肠杆菌中高效表达,纯化后的抗体具有良好的特异性,且该抗体具有生产操作简单、制备周期短和成本低等特点。
    • 吴鹏; 王英皎; 孟丽红; 高诗博; 徐军
    • 摘要: 目的:采用异硫氰酸荧光素标识Ⅳ型胶原单克隆抗体,为直接或间接荧光方法检测人体肝脏或血液中Ⅳ型胶原含量奠定基础。方法采用直接透析方法进行荧光素标识,标识后通过Sephadex G25凝胶过滤,除去多余的荧光素和其他杂质,获得标识荧光抗体纯品。经SDS-PAGE和双向免疫扩散凝胶电泳,对标识抗体进行纯度,荧光特性,抗体特异性进行检测鉴定。结果经全自动化学发光荧光图像分析仪检测SDS-PAGE电泳结果,激发波490nm,发射波510nm,在150KD处有一条黄绿荧光带,其他处未见条带出现,凝胶图像分析仪检测纯度达96%;免疫琼脂双向扩散电泳结果显示,标识抗体与未标识抗体与抗原之间均有乳白色沉淀线,且沉淀物含量一致,荧光分析仪检测标识抗体与中心孔之间有黄绿色荧光条带。结论采用本研究方法制备的荧光抗体具有纯度高、荧光性好、特异性强等特点,为临床检测人体Ⅳ型胶原含量提供便利。%Objective To lay the foundation of detecting the content of Ⅳ type collagen in human liver or blood through direct or indirect fluorescent method, Identifying monoclonal antibody of collagen type Ⅳ by using the fluorescein isothiocyanate. Methods Using a direct dialysis method to identification of fluorescein. The purity of the identifying fluorescent antibody were obtained by Sephadex G-25 gel filtration column, which remove excess fluorescein and other impurities. The purity, fluorescence characteristic and specificity of antibody were measured with SDS-PAGE and double immune diffusion gel electrophoresis. Results The results of SDS-PAGE electrophoresis were measured by fully automatic chemiluminescence fluorescence image analyzer. Eexcitation wave 490nm and emission wave 510nm. The purified of antibody were shown only one strap of yellow with green fluorescence and the molecular weight (MW) of them were estimated to be approximately 150kD, its purity was 96%, there were not any other strap. The result of double agar gel immune-diffusion test showed that both identify antibodies and not identify antibody were combination with antigen, all of product were formed milky white precipitation lines of antigen-antibody complexes with antigen. And the sediment content was consistent. There was a strap of yellow with green fluorescence in the center hole by fluoroanalyzer. Conclusion In this study, the fluorescent antibody against human collagen type Ⅳ with high purity and strong specificity was produced by separation and purification methods, which provide convenience for clinical detection of the human bodyⅣtype collagen.
    • 米立娟; 张守峰; 刘晔; 王述超; 扈荣良
    • 摘要: Four monoclonal antibodies against Rabies virus phosphoprotein were produced by immunization with Rabies virus strain CVS in BALB/c mice. After fusion, antibodies in the supernatant of the hybrid cells were respectively detected by ELISA test and western-blot method. Four hybrid cells secreting antibody binding to rabies virus , i.e. 1C9, 4B10, 2G12 and4G5 strains. The 1C9 monoclonal lgG antibody was purified by affinity chromatography method and then labeled with FITC. 501 brain samples collected from ferret badgers, bats, dogs and weasels were detected using the labeled 1C9 antibody and FITC-labeled rabies virus N protein-specific monoclonal antibody (made in our laboratory). The results showed that the labeled 1C9 antibody can detect rabies-related lyssaviruses isolated in China including Rabies virus and Irkut virus, however, the N protein-specific monoclonal antibody only can detect Rabies virus. It suggests that the labeled 1C9 antibody can be used as a rabies diagnostic reagent in China.%以灭活的狂犬病病毒CVS株细胞毒免疫BALB/c小鼠,通过间接ELISA法和Western-blot筛选获得针对磷蛋白的单克隆抗体4株:1C9、4B10、2G12、4G5,其中1C9针对氨基端保守表位。以亲和层析法纯化1C9单抗腹水,异硫氰酸荧光黄标记制备荧光抗体。以1C9磷蛋白荧光抗体与本实验室研制的狂犬病核蛋白免疫荧光抗原检测试剂盒,对本实验室收集的501份疑似狂犬病鼬獾、蝙蝠、犬和黄鼬的脑组织样品进行直接免疫荧光平行检测。结果显示,两种检测手段对基因1型狂犬病毒的检出结果完全一致,而蝙蝠源Irkut病毒仅能以磷蛋白单抗1C9检出。本研究成功获得了与我国现有不同基因型狂犬病毒良好反应的抗狂犬病磷蛋白单抗,并应用于狂犬病的直接免疫荧光检测,为狂犬病诊断提供了敏感性和可靠性良好的诊断试剂。
    • 王兵
    • 摘要: 党的十八大以后,党中央、国务院进一步要求我们重视民生问题,关心人民群众的身体健康。作为医疗部门、鉴定部门要积极响应党中央、国务院的号召,落实好关心人民群众身体健康的有关措施。新时期新形势下,感染性的疾病仍然是危害人民群众身体健康的大隐患。由于一些新的突发性感染疾病的出现,不少过去已被控制的感染性疾病又死灰复燃,造成了感染性疾病的微生物种类增多而又复杂。这些现实给新时期的临床微生物检验,诊断提出了更新更高的要求。作为医疗事故技术鉴定部门应该更加努力地在临床微生物检验诊断方面开展好探索与研究。
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