title: [Recombinant phosphoenolpyruvate carboxykinase from Mycobacterium tuberculosis for sero diagnosis].
abstract: OBJECTIVE
  To express phosphoenolpyruvate carboxykinase (PEPCK) of tuberculosis and evaluate its immune diagnostic ability.
  
  
  METHODS
  PEPCK was cloned and expressed in prokaryotic expression system. We analyzed immune response of recombinant PEPCK with Western blot and studied the immunity of it through mice immunization. We also investigated the sensitivity and specificity of immunodiagnosis of PEPCK via detecting the sera from tubercular patients by using ELISA. The results were evaluated by comparing with a Mycobacterium tuberculosis antibody Colloidal Gold Diagnostic Kit.
  
  
  RESULTS
  The recombinant protein was 72 kDa and specifically reacted with anti-BCG antibody. Specific humoral immune response was elicited after mouse immunization with PEPCK protein and the high titer specific antibody was generated (1:1280). Antibodies were detected against M. tuberculosis 17.3% in all tuberculosis patients, 32.5% in patients harboring Mycobacterium currently and 12.9% in patients without M. tuberculosis. The sensitivity and specificity of ELISA were 51.0% and 96.7%.
  
  
  CONCLUSIONS
  These results indicated that the recombinant PEPCK might be one of the suitable antigens for serodiagnosis of tuberculosis.
authors.affiliation: Department of Pathogenic Biology, Binzhou Medical University, Binzhou 256603, China. xuelianbai99@163.com
authors.forename: Xuelian
authors.initials: X
authors.lastname: Bai
authors.forename: Qingkuan
authors.initials: Q
authors.lastname: Wei
authors.forename: Jin
authors.initials: J
authors.lastname: Li
authors.forename: Gulping
authors.initials: G
authors.lastname: Li
mesh_terms: D000818:Animals; D000907:Antibodies, Bacterial / Q000097:blood; D001426:Bacterial Proteins / Q000235:genetics / Q000276:immunology*; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods*; D005260:Female; D006801:Humans; D007114:Immunization; D008297:Male; D051379:Mice; D008807:Mice, Inbred BALB C; D009169:Mycobacterium tuberculosis / Q000235:genetics / Q000276:immunology*; D019764:Phosphoenolpyruvate Carboxykinase (ATP) / Q000235:genetics / Q000276:immunology*; D011897:Random Allocation; D011994:Recombinant Proteins / Q000235:genetics / Q000276:immunology; D014376:Tuberculosis / Q000175:diagnosis* / Q000276:immunology
chemical_list: D000907:Antibodies, Bacterial; D001426:Bacterial Proteins; D011994:Recombinant Proteins; D019764:Phosphoenolpyruvate Carboxykinase (ATP)