The bromocresol green assay, but not the modified bromocresol purple assay, overestimates the serum albumin concentration in nephrotic syndrome through reaction with α2-macroglobulin.

Tsuyoshi Ueno, Satoshi Hirayama, Masami Sugihara, Takashi Miida

Annals of clinical biochemistry 2016 Jan

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The bromocresol green (BCG) assay is commonly used for measuring albumin (ALB), but is affected by α1- and α2-globulins, which are elevated in systemic inflammation. The modified bromocresol purple (mBCP) assay is another dye-binding method developed to overcome non-specific reactions. Concentrations of α2-macroglobulin, a major α2-globulin component, are increased in nephrotic syndrome (NS), but not in inflammation. There is little direct evidence that α2-macroglobulin affects BCG or mBCP assays. We measured serum albumin concentrations in 33 patients with NS and 13 reference healthy controls using BCG (ALBBCG) and mBCP (ALBmBCP) assays, and nephelometry (nALB) as a reference method. We also determined five specific proteins belonging to the α1- and α2-globulins by nephelometry. After adding purified α2-macroglobulin to albumin solutions, protein reactivity in these three assays was compared. Nephrotic syndrome patients were categorized to tertiles according to nALB concentration. In all tertiles, ALBBCG was significantly higher than nALB, especially in the severe hypoalbuminemia group, in which α2-macroglobulin was 43-49% higher. By contrast, ALBmBCP and nALB were almost identical in all three groups. The difference between ALBBCG and nALB was positively correlated with the α2-macroglobulin concentration. In vitro, when α2-macroglobulin was added to solutions containing identical albumin concentrations, α2-macroglobulin dose-dependently increased ALBBCG, but not ALBmBCP. In NS, α2-macroglobulin is a major factor for positive bias of ALBBCG, especially in patients with severe hypoalbuminemia. The mBCP assay is useful for measuring albumin concentrations in NS. © The Author(s) 2015.

Citation

Tsuyoshi Ueno, Satoshi Hirayama, Masami Sugihara, Takashi Miida. The bromocresol green assay, but not the modified bromocresol purple assay, overestimates the serum albumin concentration in nephrotic syndrome through reaction with α2-macroglobulin. Annals of clinical biochemistry. 2016 Jan;53(Pt 1):97-105