期刊文献+

cpn0585蛋白的免疫活性及血清学诊断初步评价

Preliminary Evaluation of Immunocompetence and Serological Diagnosis of Cpn0585 Protein
下载PDF
导出
摘要 目的克隆肺炎嗜衣原体包涵体膜蛋白Cpn0585基因,初步探讨其在血清学诊断中的应用价值。方法构建pGEX-6p-2/Cpn0585重组质粒,诱导表达并纯化重组蛋白,免疫BALB/c小鼠,间接酶联免疫附试验(ELISA)检测小鼠血清抗体滴度,以重组蛋白作为ELISA包被抗原检测血清中Cpn IgG抗体和检测36份Cpn-/Ct+IgG阳性参考血清。结果高效表达和纯化出一相对分子质量约为95 kDa的重组蛋白,蛋白质印迹法证明其能与人抗Cpn IgG抗体发生反应;在被免疫的BALB/c小鼠体内,特异性IgG抗体的滴度为1:12 800;检测104例临床血清标本中的IgG抗体,与以色列Savyon Diagnostics公司SeroCP^(TM)IgG ELISA诊断试剂盒的检测结果进行比较,符合率为98.3%。结论表达的Cpn0585重组蛋白具有良好的免疫活性,在Cpn的血清学诊断中具有较高的应用价值。 Objective To clone and express the Cpn0585 gene from Chlamydophila pneumonia e. and then evaluate the clinical value of the recombinant protein in serodiagnosis. Methods The pGEX-6p-2/Cpn0585 recombinant plasmid was contructed, and then expression of recombinant protein was induced. The expressed recombinant protein was identified by SDS-PAGE. BALB/c rats were immunized with the purified protein;indirect ELISA was applied to measuring its immunogenieity; and its immunoreactivity was analyzed by reacting with Cpn-specific IgG antibody in sera. Cpn IgG and Chlamydozoa trachomatis positive sera were detected with indirect ELISA. Results The recombinant protein with relative molecular weight about 95 kDa was expressed and purified effectively. Western blot analysis proved that the recombinant protein could react with human anti Cpn IgG specifically. The titer of the specific IgG antibodies in the immuned BALB/c rats was above 1:12 800. The compliance rate between the indirect ELISA test and SeroCPTM lgG ELISA kits to104 patients sera were 98. 3%. Conclusion The expressed recombinant protein of Cpn0585 from Cpn shows excellent irnmunoc-ompetence. It is suitable for preparation of serodiagnostic EIISA reagent for Cpn infection.
出处 《中南医学科学杂志》 CAS 2011年第5期524-528,共5页 Medical Science Journal of Central South China
关键词 肺炎嗜衣原体 Cpn0585 免疫活性 血清学诊断 chlaraydophila pneumoniae Cpn0585 immunocompetence serodiagnosis
  • 相关文献

参考文献20

  • 1郑江花,吴移谋,刘佳强,刘广超,陈超群.肺炎嗜衣原体蛋白酶样活性因子免疫优势区基因重组蛋白在早期诊断中的应用[J].微生物学报,2008,48(4):520-525. 被引量:4
  • 2周洲,吴移谋,刘劼,陈超群,杨玲.肺炎衣原体ompA基因VD2-VD3区重组蛋白在血清学诊断中的初步应用[J].微生物学报,2007,47(3):512-516. 被引量:6
  • 3Littman AJ,White E,Jackson LA,et al.Chlamydia pneumoniae infection and risk of lung cancer.Cancer Epidemiol. Biomarkers Prey . 2003
  • 4Ciervo A,Petrucca A,Visca P,et al.Evaluation and optimization of ELISA for detection of anti-Chlamydophila pneumoniae IgG and IgA in patients with coronary heart diseases. Journal of Microbiological Methods . 2004
  • 5Grayston J T.Chlamydiae Pneumoniaeand atherosclerosis〔J〕. Clinical Infectious Diseases . 2005
  • 6Boman J,Hammerschlag MR.Chlamydia pneumoniae and atherosclerosis: critical assessment of diagnostic methods and relevance to treatment studies. Clinical Microbiology Reviews . 2002
  • 7Campbell L A,Perez M M,Hamilton D J,et al.Detection of chlamydia pneumoniae by polymerase chain reaction. Journal of Clinical Microbiology . 1992
  • 8Apfalter P,Blasi G,Boman J,et al.Multicenter comparison trial of DNA extraction methods and PCR assays for detection of Chlamydia pneumoniae in endarterectomy specimens. Journal of Clinical Microbiology . 2001
  • 9Fainardi E,Castellazzi M,Seraceni S,et al.Under the Microscope: Focus on Chlamydia pneumoniae Infection and Multiple Sclerosis. Current Neurovascular Research . 2008
  • 10Mukhopadhyay S,Good D,Miller RD,et al.Identification of Chlamydia pneumoniae proteins in the transition from reticulate to elementary body formation. Molecular and Cellular Proteomics . 2006

二级参考文献22

  • 1周洲,吴移谋.衣原体主要外膜蛋白的研究现状[J].国外医学(微生物学分册),2005,28(2):29-31. 被引量:2
  • 2陈丽丽,吴移谋,雷达,吴志周,黄澍杰.泌尿生殖道沙眼衣原体omp1基因多态性研究[J].微生物学报,2006,46(2):214-218. 被引量:8
  • 3周洲,吴移谋,刘劼,陈超群,杨玲.肺炎衣原体ompA基因VD2-VD3区重组蛋白在血清学诊断中的初步应用[J].微生物学报,2007,47(3):512-516. 被引量:6
  • 4Hermann C,Craf K,Groh A,et al.Comparison of eleven commercial tests for Chlamydia pneunomiae-specific immunoglobulin G in asymptomatic healthy individuals.J Clin Microbiol,2002,40(5):1603-1609.
  • 5Essig A,Simnacher U,Susa M,et al.Analysis of the humoral immune rcsponse to Chlamydia pneumoniae by immunoblotting and immunopreeipitation.Clin Diagn Lab hamunol,1999,6(6):819-825.
  • 6Bennedsen M,Berthelsen L,Lind I,et al.Performance of three Microimmunofluorescence assay for detection of Chlamydia pneumoniae immunoglobulin M,G,and A antibodies.Clin Diagn Lab Immunol,2002,9(4):833-839.
  • 7Klein M,Kotz A,Bernardo K,et al.Detection of Chlamydia pneumoniae-specific antibodies binding to the VD2 and VD3 regions of the Major outer membrane protein.J Clin Microbiol,2003,41(5):1957-1962.
  • 8Read TD,Brunham RC,Shen C,et al.Genome sequence of Chlamydia trachomatis MoPn and Chlamydia pneumoniae AR39.Nucleic Acids Reseach,2000,28(6):1397-1406.
  • 9Jantos CA,Heck S,Roggendorf R,et al.Antigenic and molecular analyses of different Chlamydia pneumoniae strains.J Clin Microbiol,1997,35(3):620-623.
  • 10Bosnic AM, Piper JM, et al. Human antibody responses to a Chlamydia-secreted protease factor.Infect Immun, 2004, 72 (12): 7164-7171.

共引文献8

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