Development of a sensitive enzyme-linked immunosorbent assay for measurement of DNase I in human serum

Clin Chim Acta. 2009 May;403(1-2):219-22. doi: 10.1016/j.cca.2009.03.005. Epub 2009 Mar 21.

Abstract

Background: Serum deoxyribonuclease I (DNase I) activity was reported to increase in the early phase after onset of acute myocardial infarction (AMI). Up to now, DNase I activity has been quantified by the single radial enzyme diffusion (SRED) method, which unfortunately requires a long incubation time. Therefore it is necessary to develop another assay suitable for measurement of serum DNase I concentrations in a clinical setting.

Methods: A sandwich ELISA was established for measurement of DNase I protein using a polyclonal antibody directed against DNase I protein and a biotinylated monoclonal for subsequent detection. Concentrations of serum DNase I protein were measured in healthy individuals and patients with AMI.

Results: This method was as precise as SRED, and took less time than SRED. A significant correlation was observed between DNase I concentration and enzyme activity (r=0.839; P<0.001). The average of serum DNase I in AMI patients within 0-12 h of chest pain was significantly higher than that in healthy individuals (P<0.001), and decreased with time.

Conclusions: We have developed a sensitive ELISA capable of measuring DNase I protein concentrations. This method may be a useful alternative to SRED as an aid to diagnosis of AMI based on the serum DNase I level.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Adult
  • Antibodies / immunology
  • Antibody Specificity
  • Deoxyribonuclease I / blood*
  • Deoxyribonuclease I / immunology
  • Enzyme-Linked Immunosorbent Assay / methods*
  • Female
  • Humans
  • Male
  • Middle Aged
  • Myocardial Infarction / blood
  • Myocardial Infarction / enzymology
  • Sensitivity and Specificity
  • Time Factors

Substances

  • Antibodies
  • Deoxyribonuclease I