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Updated: Jan 14, 2026
Detection of Inflammasome Activation and Pyroptotic Cell Death in Murine Bone Marrow-derived Macrophages
Published on: May 21, 2018
A semi-automated ASC speck assay to evaluate pyrin inflammasome activation
Pei Dai1,2,3, Oliver Skinner4,5, Xufeng Lin6
1Precision Immunology Program Garvan Institute of Medical Research Sydney NSW Australia.
Objective:
To develop a rapid functional assay to validate variants of uncertain significance (VUS) in the MEFV gene.
Methods:
Overactivity of the pyrin inflammasome pathway and ASC speck oligomerisation in response to stimulation with low concentrations of Clostridium difficile toxin A was directly visualised by immunofluorescence microscopy. A semi-automated algorithm was developed to count cells and ASC specks.
Results:
The semi-automated ASC speck assay is able to discriminate between healthy controls and patients with familial Mediterranean fever (FMF) and pyrin inflammasome overactivity with high sensitivity. It is also able to discriminate pyrin inflammasome overactivity from other autoinflammatory disease controls with high specificity.
Conclusion:
The semi-automated ASC speck assay may be a useful test to functionally validate VUS in the MEFV gene and screen for pyrin inflammasome overactivity.
