title: [Establishment of enzyme-linked immunosorbent assay (ELISA) for measuring human urinary uromodulin and application of the method in patients with IgA nephropathy].
abstract: OBJECTIVE
  To establish a method of enzyme-linked immunosorbent assay (ELISA) to measure urinary uromodulin and explore the urinary uromudulin  level in IgA nephropathy.
  
  
  METHODS
  The rabbit anti-human uromodulin polyclonal antibodies were coated on plates to capture uromodulin and the  mouse anti-human uromodulin monoclonal antibody was used as detecting antibody to set up ELISA procedure. The precision and repeatability of this  ELISA method were evaluated, and then this method was compared with the commercialized Tamm-Horsfall Glycoprotein ELISA Kit by examining urinary  uromodulin levels in 55 individuals. Finally, the urinary uromodulin level in 166 IgA nephropathy patients were detected as well as 48 normal controls with  this established method.
  
  
  RESULTS
  The detecting range of uromodulin was 0.78-12.5 μg/L by this method. The coefficient of variation within-run was 7.5%,  and between-run of coefficient of variation was 7.9%. Correlation of this method and comercialized kit was good (r=0.615, P<0.001). The urinary  uromodulin/urinary creatinine ratio in IgA nephropathy was significantly lower than that in normal controls.
  
  
  CONCLUSIONS
  The established ELISA method is  sensitive and repeatable, and can be used in further studies.
authors.affiliation: Renal Division, Peking University First Hospital; Peking University Institute of Nephrology; Key Laboratory of Renal Disease, Ministry of Health of China; Key Lab of Chronic Kidney Disease Prevention and Treatment, Ministry of Education; Beijing 100034, China.
authors.forename: Ying
authors.initials: Y
authors.lastname: Liu
authors.forename: Yu-qing
authors.initials: YQ
authors.lastname: Chen
authors.forename: Jing-jing
authors.initials: JJ
authors.lastname: Zhou
authors.forename: Jia
authors.initials: J
authors.lastname: Han
authors.forename: Yu
authors.initials: Y
authors.lastname: Liang
authors.forename: Xue-ying
authors.initials: XY
authors.lastname: Li
authors.forename: Hong
authors.initials: H
authors.lastname: Zhang
mesh_terms: D000328:Adult; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods*; D005260:Female; D005922:Glomerulonephritis, IGA / Q000652:urine*; D006801:Humans; D008297:Male; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D058949:Uromodulin / Q000652:urine*
chemical_list: D058949:Uromodulin