A Rapid Sputum-based Lateral Flow Assay for Airway Eosinophilia using an RNA-cleaving DNAzyme Selected for Eosinophil Peroxidase |
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| Authors: | Dr. M. Monsur Ali Prof. Dr. Manali Mukherjee Katherine Radford Zil Patel Prof. Dr. Alfredo Capretta Prof. Dr. Parameswaran Nair Prof. Dr. John D. Brennan |
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| Affiliation: | 1. Biointerfaces Institute, McMaster University, 1280 Main Street West, L8S 4K1 Hamilton, ON, Canada;2. Division of Respirology, McMaster University, Firestone Institute of Respiratory Health at St. Joseph's Health Care, L8N 4A6 Hamilton, ON, Canada |
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| Abstract: | The first protein-binding allosteric RNA-cleaving DNAzyme (RCD) obtained by direct in vitro selection against eosinophil peroxidase (EPX), a validated marker for airway eosinophilia, is described. The RCD has nanomolar affinity for EPX, shows high selectivity against related peroxidases and other eosinophil proteins, and is resistant to degradation by mammalian nucleases. An optimized RCD was used to develop both fluorescence and lateral flow assays, which were evaluated using 38 minimally processed patient sputum samples (23 non-eosinophilic, 15 eosinophilic), producing a clinical sensitivity of 100 % and specificity of 96 %. This RCD-based lateral flow assay should allow for rapid evaluation of airway eosinophilia as an aid for guiding asthma therapy. |
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| Keywords: | Asthma DNAzyme Eosinophilic Peroxidase Lateral Flow Assay Sputum |
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