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Annals of Clinical & Laboratory Science 35:149-154 (2005)
© 2005 Association of Clinical Scientists

Microplate ELISA for Urine Microalbumin: Reference Values and Results in Patients with Type 2 Diabetes and Cardiovascular Disease

Tsu-Lan Wu1, Pi-Yueh Chang1, Chia-Chi Li1, Kuo-Chien Tsao1, Chien-Feng Sun1 and James T. Wu2
1 Department of Pathology, Chang Gung Memorial Hospital, Taipei, Taiwan; 2 ARUP Laboratories, University of Utah Health Sciences Center, Salt Lake City, Utah

Address correspondence to James T. Wu, Ph.D., ARUP Laboratories, 500 Chipeta Way, Salt Lake City, UT 84108, USA; tel 801 583 2787; fax 801 584 5207; e-mail wuj{at}aruplab.com.

An ELISA for urine microalbumin using microtiter plates has been developed. The assay uses polyclonal anti-human albumin antibody for coating the microtiter plates and the same antibody conjugated with horseradish peroxidase for detection. The assay sensitivity is 1.6 µg/ml. Results by this in-house ELISA show good correlation (r = 0.99) with those obtained by a commercial assay using the Behring BNII autoanalyzer. Within-day and between-day CVs are 10%. R eference values for microalbumin in 769 urine specimens from healthy Chinese subjects were higher in women than men and higher in subjects 50 yr than in those <50 yr of age. Elevated mean concentrations of urine microalbumin were observed in patients with type 2 diabetes and CVD. This in-house ELISA is simple, sensitive, precise, and especially suited for laboratories without expensive autoanalyzers.

Keywords: enzyme-linked immunosorbent assay, type 2 diabetes, cardiovascular disease, microtiter plate assay, urine microalbumin

Abbreviations: BSA-PBS: phosphate buffer, 0.01 mol/L, pH 7.2, containing bovine serum albumin, 10 g/L; BNII, Behring Nephelometric Analyzer II; CRP, C-reactive protein; CVD, cardiovascular disease; ELISA, enzyme-linked immunosorbent assay; HRP, horse radish peroxidase; IL-6, interleukin-6; TNF-{alpha}, tumor necrosis factor-alpha; PBS, phosphate buffer, 0.1 mol/L, pH 7.2, containing NaCl, 0.14 mol/L, K-Blue, tetramethylbenzidine.




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