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Automation of o-dianisidine assay for ceruloplasmin activity analyses: usefulness of investigation in Wilson’s disease and in hepatic encephalopathy

  • Neurology and Preclinical Neurological Studies - Original Article
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Abstract

Ceruloplasmin (Cp) is a serum ferroxidase that plays an essential role in iron metabolism. It is routinely tested by immunoturbidimetric assays that quantify the concentration of the protein both in its active and inactive forms. Cp activity is generally analyzed manually; the process is time-consuming, has a limited repeatability, and is not suitable for a clinical setting. To overcome these inconveniences, we have set the automation of the o-dianisidine Cp activity assay on a Cobas Mira Plus apparatus. The automation was rapid and repeatable, and the data were provided in terms of IU/L. The assay was adapted for human sera and showed a good precision [coefficient of variation (CV) 3.7 %] and low limit of detection (LoD 11.58 IU/L). The simultaneous analysis of Cp concentration and activity in the same run allowed us to calculate the Cp-specific activity that provides a better index of the overall Cp status. To test the usefulness of this automation, we tested this assay on 104 healthy volunteers and 36 patients with Wilson’s disease, hepatic encephalopathy, and chronic liver disease. Cp activity and specific activity distinguished better patients between groups with respect to Cp concentration alone, and providing support for the clinical investigation of neurological diseases in which liver failure is one of the clinical hallmarks.

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Abbreviations

Cp:

Ceruloplasmin

eCp:

Enzymatic Cp

iCp:

Immunologic Cp

CTRL:

Healthy volunteers

WD:

Wilson’s disease

CLD:

Chronic liver disease

ENC:

Hepatic encephalopathy

HCV:

Hepatitis C virus

CV:

Coefficient of variation

LoD:

Limit of detection

LoB:

Limit of blank

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Acknowledgments

The authors thank Dr.Vernieri F., Dr. Altamura, C. (Department of Neurology) and Dr. Vespasiani Gentilucci U., Dr. Picardi A. (Department of Clinical Medicine and Hepatology) of University Campus Bio-Medico of Rome, Italy, for providing the samples. This study was partially supported by the Grant: Italian Health Department: “Profilo Biologico e Genetico della Disfunzione dei Metalli nella Malattia di Alzheimer e nel ‘Mild Cognitive Impairment” [RF 2006 conv.58].

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Authors and Affiliations

  1. Don Carlo Gnocchi Foundation ONLUS, P.le Morandi 6, 20121, Milan, Italy

    Mariacristina Siotto

  2. Medical Statistics and Information Technology, Fatebenefratelli Foundation for Health Research and Education, AFaR Division, Rome, Italy

    Patrizio Pasqualetti

  3. Unit of Clinical and Molecular Epidemiology, IRCCS “San Raffaele Pisana”, Rome, Italy

    Patrizio Pasqualetti

  4. Department of Neurology, University “Campus Bio-medico”, Via Alvaro del Portillo 21, 00128, Rome, Italy

    Massimo Marano

  5. Fatebenefratelli Foundation for Health Research and Education, AFaR Division, Rome, Italy

    Rosanna Squitti

  6. Laboratorio di Neurodegenerazione, IRCCS “San Raffaele Pisana”, Rome, Italy

    Rosanna Squitti

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  1. Mariacristina Siotto
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  2. Patrizio Pasqualetti
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  3. Massimo Marano
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  4. Rosanna Squitti
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Correspondence to Mariacristina Siotto.

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Siotto, M., Pasqualetti, P., Marano, M. et al. Automation of o-dianisidine assay for ceruloplasmin activity analyses: usefulness of investigation in Wilson’s disease and in hepatic encephalopathy. J Neural Transm 121, 1281–1286 (2014). https://doi.org/10.1007/s00702-014-1196-0

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  • DOI: https://doi.org/10.1007/s00702-014-1196-0

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