期刊文献+

牛结核病抗体胶体金快速检测技术的建立和应用 被引量:26

Establishment and Preliminary Application of Immunochromatographic Strips for Rapid Detection of Antibodies against M.bovis
下载PDF
导出
摘要 为了建立一种快速检测牛分支杆菌抗体的新方法用于诊断牛结核病,利用胶体金免疫层析技术原理,用原核诱导表达的牛分支杆菌抗原蛋白MPB83和MPB70分别作为胶体金标记抗原和检测线上的捕获抗原,制备牛结核抗体检测试纸条。结果表明,粒径为40 nm的胶体金制备的试纸条敏感性最高,胶体金最佳标记pH为6.0,MPB83抗原最适标记量为每毫升胶体金6.5μg,MPB70抗原的最适包被浓度为3.0 mg/mL,抗MPB83蛋白IgG的最佳包被浓度为2.5 mg/mL,交叉试验证明试纸条不与牛的其他非相关疾病的阳性血清反应,具有较高的特异性。比较试验证明其敏感性显著高于韩国进口试纸条。在上述试验条件下生产了一批胶体金试纸条进行临床样品检测,并与细菌分离培养、结核菌素皮内变态反应(TST)和韩国试纸条比较。本试纸条与牛分支杆菌分离培养的符合率为85%,与TST的符合率为79.73%,与韩国试纸条的符合率为98.75%。快速检测牛结核抗体的免疫层析试纸条具有敏感、特异、简便、快速的特点,适用于对牛结核病进行普查和检疫,也可作为TST的辅助诊断方法,在牛结核病根除计划中具有良好的应用前景。 To develop a new method to rapidly detect antibodies against Mycobacteriurn boris for diagnosis of bovine tuberculosis(bTB). Strips were prepared with the principle of colloidal gold immunochromatography assay (GICA) with the two recombinant Mycobacteriurn boris antigens MPB83 and MPB70. MPB83 was labeled by colloidal gold and MPB70 was used as the test capture reagent to detect the corresponding antibodies. The highest sensitivity was found with strips prepared by 40 nm particles of colloidal gold. The optimal labeling pH of colloidal gold solution was 6.0. The best amount of MPB83 for colloidal gold labelling was 6.5 g per ml. The optimal concentration of capture reagent MPB70 and control coating protein anti-MPB83 IgG was 3.0mg/mL and 2.5mg/mL, respectively. The strips did not react with positive sera of other unrelated bovine diseases and displayed high specificity. Comparison test showed that the strips had a higher sensitivity than a commercial MPB70 test strips from Korea. A batch of colloidal gold test strips were manufactured to detect clinical samples, in parallel with culture of M. boris, tuberculin skin testing (TST) and test strips from Korea. The agreement ratios were 85% ,79.73% and 98.75% ,respectively, between this strip with M. boris isolation, TST, and the Korea MPB70 strip. This novel colloidal gold test strip for bTB antibody detection is sensitive, specific, convenient to be used, rapid to get results. It will be a good method in bTB survey used alone or as a supplementary diagnostic method for TST in bTB eradication program.
出处 《动物医学进展》 CSCD 2007年第7期17-24,共8页 Progress In Veterinary Medicine
基金 国家"十一五"科技攻关奶业重大专项课题(2006BAD04A05) 武汉市科技攻关课题(20066002056)
关键词 免疫层析 牛分支杆菌 牛结核 抗体检测 immunochromatography Mycobacterium boris bovine tuberculosis antibody detection
  • 相关文献

参考文献16

  • 1王忠仁.牛结核病与人结核病的相互关系[J].中国防痨杂志,2005,27(2):123-125. 被引量:35
  • 2Gordejo R F J,Vermeersch J P.Towards eradication of bovine tuberculosis in the European Union[J].Vet Microbiol,2006,112(2-4):101-109.
  • 3Domenech R.Human Mycobacterium bovis infection in the United Kingdom:Incidence,risks,control measures and review of the zoonotic aspects of bovine tuberculosis[J].Tuberculosis (Edinb),2006,86 (2):77-109.
  • 4杨卫冲,焦新安.牛结核病诊断技术的研究进展[J].中国人兽共患病杂志,2004,20(12):1090-1093. 被引量:31
  • 5Rhodes S G,Widen D,Buddle B M,et al.Antigen specificity in experimental bovine tuberculosis[J].Infect Immun,2000,68(5):2573-2578.
  • 6Shiff C J,Premji Z,Minjas J N.The rapid manual ParaSight-F test.A new diagnostic tool for Plasmodium falciparum infection[J].Trans Rsoc Trop Med Hyg,1993,87(6):646-648.
  • 7Greenwald R,Esfandiari J,Lesellier S,et al.Improved serodetection of Mycobacterium bovis infection in badgers (Meles meles) using multiantigen test formats[J].Diag Microbiol Infect Dis,2003,46:197-203.
  • 8Wiker H G,Lyashchenko K P,Aksoy M A,et al.Immunochemical characterization of the MPB70/80 andMPB83 proteins of Mycobacterium bovis[J].Infection and Immunity,1998,66(4):1445-1452.
  • 9王文勇,李玉松,赵一岭.一种快速、稳定制备胶体金的新方法[J].细胞与分子免疫学杂志,1997,13(A01):18-19. 被引量:30
  • 10霍峰,惠观涛.应用琼脂扩散法检测小鹅瘟抗原抗体[J].中国兽医杂志,2000,26(2):21-22. 被引量:10

二级参考文献34

  • 1王忠仁.结核病传染与预防对策(现代实用结核病系统讲座第一讲)[J].中华结核和呼吸杂志,1994,17(5):267-269. 被引量:23
  • 2蔡文琴,实用免疫细胞化学与核酸分子杂交技术,1994年,98页
  • 3李玉松,中国组织化学与细胞化学杂志,1993年,2卷,1期,91页
  • 4Reilly L M ,Daborn C J. The epidemiology of Mycobacterium bovis infection in animals and man: A review[J]. Tubercle and Lung Diseasis, 1995,76:1~46.
  • 5Wedlock DN,Skinner MA, Lisle GW, et al. Review:Control of Mycobacterium bovis infections and the risk to human populations[J]. Microbes Infect, 2002,4(4):471-480.
  • 6黄纪铨.牛结核病[J].福建畜牧兽医,2000,22(6):39-39.
  • 7朱长光 李志军 李宏伟.北京市某奶牛场检出结核、副结核阳性牛[J].中国动物检疫,1999,16(4):19-20.
  • 8Surujballi OP,Romanowska A, Sugden EA, et al. A fluorescence polarization assay for the detection of antibodies to Mycobacterium bovis in cattle sera[J]. Vet Microbiol,2002,87(2): 149-157.
  • 9Silva E. Evaluation of an enzyme-linked immunosorbent assay in the diagnosis of bovine tuberculosis[J]. Vet Microbiol,2001,78(2): 111-117.
  • 10Amadori M, Lyashchenko KP, Gennaro ML, et al. Use of recombinant proteins in antibody tests for bovine tuberculosis[J]. Vet Microbiol, 2002, 85(4): 379-389.

共引文献98

同被引文献327

引证文献26

二级引证文献145

相关作者

内容加载中请稍等...

相关机构

内容加载中请稍等...

相关主题

内容加载中请稍等...

浏览历史

内容加载中请稍等...
;
使用帮助 返回顶部