Chemiluminescent immunoassay (CLIA) for urinary albumin |
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Authors: | F Javier Bagazgoitia J Luis Garcia J M Ibarra R Patio Carlos Diquez Ian Weeks J Stuart Woodhead |
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Institution: | F. Javier Bagazgoitia,J. Luis Garcia,J. M. Ibarra,R. Patiño,Carlos Diéquez,Ian Weeks,J. Stuart Woodhead |
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Abstract: | A simple chemiluminescent immunoassay (CLIA) for urinary albumin has been developed based on the use of a chemiluminescent acridinium ester-labelled human albumin and a commercially available antiserum. It includes two incubation steps and a second polyethylene glycol-assisted antibody separation. The sensitivity of detection is 0.016 mg/l, the assay working range is 0.1-5 mg/l, and the inter-assay CVs are ≤ 15%. Using 10? and 50-fold sample dilutions in assay buffer, a wide working range (1-250 mg/l) is obtained covering normal and pathological conditions. Timed overnight urine samples (bed rest conditions) were collected on three consecutive days for each patient. Albumin excretion rate (AER) was 4.7 ± 2.7 μg/min (x ± SD), range 1-15.9 μg/min in 36 healthy subjects (17♂, 19♀, ages 4-56 years), with day-to-day variations of 28.5 ± 20% (x ± SD), range 3.3-76.1%. The use of an acridinium ester as a chemiluminescent (CL) label overcomes the disadvantages of short shelf-life and health and safety hazards associated with radioisotopes. Results compare favourably with those obtained using a commercially available RIA kit. |
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Keywords: | Diabetes albuminuria chemiluminescent immunoassay acridinium ester |
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