title: [Microplate chemiluminescence enzyme immunoassay for the quantitative analysis of tissue inhibitor of metalloproteinases I].
abstract: OBJECTIVE
  To establish microplate chemiluminescence enzyme immunoassay (CLEIA) for quantitative analysis of tissue inhibitor of metalloproteinases I (TIMP I) in human serum.
  
  
  METHODS
  A sandwich reaction was preformed with horseradish peroxidase(HRP) labeled monoclonal antibody of TIMP I as the catalytic enzyme and the H2O2-luminol as the luminescence reagent. Several physical and chemical parameters were studied and optimized such as immunoreaction conditions, the dilution ratio of TIMP I-HRP, luminescence reaction time and so on. In order to evaluate the method, recovery test, heat stabilization test and comparison test were carried out.
  
  
  RESULTS
  The linear range was 0. 2-12 ng/ml with r = 0.996. The detection limit was 0.12 ng/ml. Inter-assay and intra-assay RSD were both less than 10%. The recoveries of three different spiked concentration samples were 100.6%, 96.5% and 106.5%. After stored at 4 degrees C and 37 degrees C for 3, 5, 7 days, the analysis showed correlation coefficient higher than 0. 998 and RSD lower than 6%. The detected results with CLEIA closely corresponded to those with imported ELISA in 60 patients sera with liver fibrosis.
  
  
  CONCLUSIONS
  Established CLEIA for quantity determination of serum TIMP I has high accuracy, sensitivity and repeatability.
authors.affiliation: Medical University of PLA, Beijing 100853, China.
authors.forename: Jun
authors.initials: J
authors.lastname: Hou
authors.affiliation: Medical University of PLA, Beijing 100853, China.
authors.forename: Jian
authors.initials: J
authors.lastname: Zhang
authors.affiliation: Medical University of PLA, Beijing 100853, China.
authors.forename: Jia
authors.initials: J
authors.lastname: Liu
authors.affiliation: Medical University of PLA, Beijing 100853, China.
authors.forename: Jing
authors.initials: J
authors.lastname: Zhao
authors.affiliation: Medical University of PLA, Beijing 100853, China.
authors.forename: Jing-Xia
authors.initials: JX
authors.lastname: Guo
authors.affiliation: Medical University of PLA, Beijing 100853, China.
authors.forename: Lin
authors.initials: L
authors.lastname: Cheng
authors.affiliation: Medical University of PLA, Beijing 100853, China.
authors.forename: Jun
authors.initials: J
authors.lastname: Xu
authors.affiliation: Medical University of PLA, Beijing 100853, China.
authors.forename: Ai-Xia
authors.initials: AX
authors.lastname: Liu
authors.affiliation: Medical University of PLA, Beijing 100853, China.
authors.forename: Yong-Ji
authors.initials: YJ
authors.lastname: Song
authors.affiliation: Medical University of PLA, Beijing 100853, China.
authors.forename: Bo-An
authors.initials: BA
authors.lastname: Li
authors.affiliation: Medical University of PLA, Beijing 100853, China.
authors.forename: Hong-Shan
authors.initials: HS
authors.lastname: Wei
mesh_terms: D000328:Adult; D000368:Aged; D005260:Female; D006801:Humans; D007124:Immunoenzyme Techniques / Q000295:instrumentation / Q000379:methods*; D008103:Liver Cirrhosis / Q000097:blood* / Q000175:diagnosis / Q000201:enzymology*; D008163:Luminescent Measurements / Q000295:instrumentation / Q000379:methods*; D008297:Male; D008875:Middle Aged; D019715:Tissue Inhibitor of Metalloproteinase-1 / Q000097:blood*
chemical_list: C560822:TIMP1 protein, human; D019715:Tissue Inhibitor of Metalloproteinase-1