期刊文献+

胰岛素瘤相关蛋白-2自身抗体酶联免疫吸附测定方法的建立 被引量:1

Establishment of Enzyme-linked Immunosorbent Assay for Insulinnoma-associated Protein-2 Autoant ibodies.
下载PDF
导出
摘要 目的:建立糖尿病患者血清中胰岛素瘤相关蛋白鄄2(IA鄄2)自身抗体的酶联免疫吸附测定(ELISA)方法。方法:以链亲和素预包被的聚苯乙烯微量滴定板为固相载体,原核表达的生物素酰化IA鄄2胞内段融合蛋白为抗原,辣根过氧化物酶标记的山羊抗人IgG为二抗,2,2’鄄连氮鄄双鄄[3鄄乙基苯并噻唑啉磺酸](ABTS)为显色底物建立ELISA。结果:本方法的平均批内变异系数为8.8%,批间变异系数为7.7%。用该方法和Roche公司生产的“DiapletsAnti鄄IA鄄2”IA鄄2自身抗体酶联免疫试剂盒进行对比,共检测218例糖尿病患者,以Roche试剂盒的检测结果为准,本方法的诊断灵敏度为92.1%,特异度为98.9%。结论:本研究建立的IA鄄2自身抗体ELISA法有较好的灵敏度、特异度和精密度,且操作简单方便,有较广阔的应用前景。 Objectiv e: To establish enzyme-linked immunosorbent assay(ELISA)for insulinnoma-associate d protein-2(IA-2)au-toantibodies in serum of patients with diabetes mellitu s.Methods:The E.coli-derived biotinylated intracytoplasmic part of IA-2fusion protein was used as antigen and it was captured on streptavidin pre-coated poly styrene microtiter plates.The horseradish peroxidase(HRP)labeled goat anti-huma n-IgG was used as secondary antibody.2,2'-azino-di-Establishment of Enzyme-linked Immunosorbent Assay for Insulinnoma-associated Protein-2 Autoant ibodies.$$$$ JIA Xiujuan,XIE Xiaoyan,ZHANG Di,et al.Ruijin Hospital,Shanghai Institut e of Endocrinology,Shanghai Second Medical University,Shanghai(200025)Objectiv e: To establish enzyme-linked immunosorbent assay(ELISA)for insulinnoma-associate d protein-2(IA-2)au-toantibodies in serum of patients with diabetes mellitu s.Methods:The E.coli-derived biotinylated intracytoplasmic part of IA-2fusion protein was used as antigen and it was captured on streptavidin pre-coated poly styrene microtiter plates.The horseradish peroxidase(HRP)labeled goat anti-huma n-IgG was used as secondary antibody.2,2'-azino-di-[3-ethylbenzthia-zoline sulfonatel(ABTS)solution was used as substrate for HRP.Results:The overall intr a-assay coefficient of variation(CV)was8.8%and the inter-assay CV was7.7%.Our kit was compared with'Diaplets anti-IA-2'IA-2autoantibodies ELISA kit produ ced by Roche company.A total of218patients with diabetes mellitus were detected. When the Roche kit was used as standard,the diagnostic sensitivity and specifici ty of our kit were92.1%and98.9%respectively.Conclu-sions:The method of ELISA sh ows good sensitivity,specificity and precision in detecting IA-2autoantibody,al so it is con-venient to be served as a routine use.
出处 《诊断学理论与实践》 2003年第2期110-112,共3页 Journal of Diagnostics Concepts & Practice
基金 上海市卫生局科技发展基金(00418) 上海市高校自然科学研究基金(2000B16)
关键词 胰岛素瘤相关蛋白-2 自身抗体 酶联免疫吸附测定 糖尿病 Insulinnoma-assoc iated protein-2Autoantibodies Diabetes mellitus ELISA
  • 相关文献

参考文献1

二级参考文献9

  • 1Bonifacio E, Lampasona V, Bingley PJ. IA-2 (islet cell antigen 512) is the primary target of humoral autoimmunity against type 1 diabetes-associated tyrosine phosphatase autoantigen [J]. J Immunol, 1998, 161:2648-2654.
  • 2Masuda M, Powell M, Chen S, et al. Autoantibodies to IA-2 in insulin-dependent diabetes mellitus: measurements with a new immunoprecipitation assay [J]. Clinica Chimica Acta, 2000, 291:53-66.
  • 3Lobner K, Khoo-Morgenthaler UY, Seissler J, et al. Detection of autoantibodies to the diabetes-associated antigen IA-2 by a sensitive enzyme-linked immunosorbent assay [J]. Horm Metab Res, 1999, 31:686-691.
  • 4Balaji M, Shtauvere BA, Balaji V, et al. Women diagnosed with gestational diabetes mellitus do not carry antibodies against minor islet cell antigens [J]. Ann N Y Acad Sci, 2002, 958:281-284.
  • 5Grasso YZ, Reddy SK, Rosenfeld CR, et al. Autoantibodies to IA-2 and GAD65 in patients with type 2 diabetes mellitus of varied duration: prevalence and correlation with clinical features [J]. Endocr Pract, 2001, 7 (5):339-345.
  • 6Hawkes CJ, Schloot NC, Marks J, et al. T-cell lines reactive to an immunodominant epitope of the tyrosine phosphatase like autoantigen IA-2 in type 1 diabetes [J]. Diabetes, 2000, 49 (3): 356-366.
  • 7Pastore MR, Balini A, Bazzigaluppi E, et al. Two-step islet autoantibody screening for risk assessment of type 1 diabetes in relatives [J]. Diabetes Care, 1998, 21: 1445-1450.
  • 8Kolm LV, Berlo S, Bonifacio E, et al. Human monoclonal antibodies isolated from type 1 diabetes patients define multiple epitopes in the protein tyrosine phosphatase-like IA-2 antigen [J]. J Immunol, 2000, 165 (8):4676-4684.
  • 9Verge CF, Stenger D, Bonifacio E, et al. Combined use of autoantibodies (IA-2 autoantibodies, GAD autoantibody, insulin autoantibody, cytoplasmic islet cell antibodies) in type 1 diabetes: combinatorial islet autoantibody workshop [J]. Diabetes, 1998, 47: 1857-1866.

共引文献1

同被引文献8

  • 1Bambra CS.Anti-trophectoderm vaccines:rationale and methods used for antigen identification and selection[J].Scand J Immunol,1992,11(Suppl):131
  • 2Frank HG,Bose P,Albieri-Borges A,et al.Evaluation of fusogenic trophoblast surface epitopes as targets for immune contraception[J].Contraception,2005,71(4):282
  • 3Del Guercio MF,Alexander J,Kubo RT,et al.Potent immunogenic short linear peptide constructs composed of B cell epitopes and Pan DR T helper epitopes (PADRE) for antibody responses in vivo[J].Vaccine,1997,15(4):441
  • 4Benirschke K,Kaufmann P.Pathology of the human placenta[M].4th ed.New York:Springer Verlag,2000.126
  • 5Das M,Xu B,Lin L,et al.Phosphatidylserine efflux and intercellular fusion in a BeWo model of human villous cytotrophoblast[J].Placenta,2004,25(5):396
  • 6Finn OJ,Gendler SJ.Mimic 1 of MUC1[J].Nat Biotechnol,1998,26(3):236
  • 7何畏,史常旭,梁志清,李玉清.多价精子抗原表位多肽-免疫刺激复合物的抗生育作用研究[J].重庆医学,2002,31(7):555-556. 被引量:6
  • 8Waiyan Candy Ng,冯明钊.T辅助细胞在疫苗研制中的作用(英文)[J].热带医学杂志,2002,2(3):207-214. 被引量:2

引证文献1

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