Development of an enhanced chemiluminescence immunoassay (CLIA) for detecting urinary albumin

• Published in: Molecular biology reports 2012-12, Vol.39 (12), p.10851-10858
• Main Authors: Meibodi, Elham Sadat Aghaei, Azizi, Maedeh Darziani, Paknejad, Malieh, Larijani, Bagher, Omidfar, Kobra
• Format: Article
• Language: English
• Subjects: Albuminuria - urine; Animal Anatomy; Animal Biochemistry; Biomedical and Life Sciences; Calibration; Histology; Humans; Immunoassay; Immunoassay - methods; Life Sciences; Luminescent Measurements - methods; Molecular biology; Morphology; Nephelometry and Turbidimetry; Proteins; Reagent Kits, Diagnostic; Regression Analysis; Sensitivity and Specificity; Serum Albumin - analysis; Specific Gravity; Urinalysis
• ISSN: 0301-4851; 1573-4978
• DOI: 10.1007/s11033-012-1981-5

In the present study, a sensitive and competitive chemiluminescence immunoassay (CLIA) was developed in order to detect human serum albumin (HSA) in urine specimen. The method utilizes a home-made monoclonal anti-albumin antibody conjugated to horseradish peroxidase enzyme (mAb-HRP). Sensitivity, specificity and linearity of the assay were evaluated. According to the results, the proper concentration of HSA and mAb-HRP conjugates was 800 ng/100 μl and 1:200 respectively. In optimal conditions, this method could detect HSA in a high linear range of 10–200 μg ml −1 with the low detection limit of 0.025 μg ml −1 . No evidence of interference with presence of probable substances in the urine samples indicated its high specificity and selectivity. Moreover, high reproducibility as well as high sensitivity and specificity of the test were confirmed using diabetic and non-diabetic samples. Significant concordance was observed between CLIA and immunoturbidimetry assay regarding detection of HSA. The results of the present study can be considered in accordance with the current demands such as reliability, accuracy, convenience and high speed of performance for a precise protein detection method. Furthermore, it may be regarded as a more rapid, simpler and cheaper alternative compared to other sophisticated assays.