Patent ID: 8399208

Claim:
A Specific Immunological Extraction Followed by Enzymatic Detection (SIEFED) method for measuring the activation status of neutrophil cells in a biological sample obtained from a mammal, the sample containing the neutrophil cells and/or enzyme released by the neutrophil cells, which method specifically measures the active enzyme content only, the content being correlated with the neutrophil cell activation status, the method comprising the steps of: immunocapturing the enzyme released by the neutrophil cells present in the biological sample by contacting the biological sample with a neutrophil enzyme specific polyclonal or monoclonal antibody immobilized on a solid support; washing to remove any components that can interfere with the measurement of the enzyme activity; adding a specific substrate that is transformed by neutrophil active enzyme into a detectable fluorometric reaction product that generates a fluorescence signal; adding an effective amount of nitrite to the reaction medium to enhance the generated fluorescent signal; and detecting and/or measuring the enzymatic activity of the immunocaptured neutrophil enzyme present which indicates the activation status of neutrophil cells in the biological sample.