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A Single Multiplex CytoBas Assay Incorporating Eight Major Components for Accurate Detection of Allergen
Lin Hsin1, Mark Hew2,3, Pei Mun Aui1
1Department of Immunology, School of Translational Medicine, Monash University, Melbourne, Victoria, Australia.
Background:
Allergic rhinitis and asthma can be triggered by a variety of aeroallergens, including house dust mites (HDM), tree and grass pollen, and household pets. Identification of the relevant allergen is critical for lifestyle changes and treatments, including allergen immunotherapy. We here assessed the diagnostic performance and clinical utility of a single flow cytometry staining of basophils with major aeroallergen components (AeroDiff CytoBas).
Methods:
In 156 atopic patients with allergic rhinitis/asthma and 21 non-atopic individuals, allergen-specific IgE levels were determined by ImmunoCAP, and component-specific IgE by ELISA. PBMCs were analyzed by flow cytometry with basophil markers and eight fluorochrome-conjugated allergen component tetramers.
Results:
Patients were stratified for sensitization to each of the four allergens. Allergen-component staining in a single multiplex CytoBas assay and component-specific IgE serology performed similarly for Der p 2, Lol p 1, Fel d 1, and Can f 1 (ROC AUC: 0.76-0.97 vs. 0.73-0.93). CytoBas had greater diagnostic accuracy than component-specific IgE serology (p < 0.001) for HDM sensitization using Der f 1 or Der p 1, and grass pollen using Lol p 5 or Phl p 1. Furthermore, the combined evaluation of Der p 1 and Der p 2 with CytoBas was 96.3% sensitive and 90.7% specific for HDM sensitization. The combined evaluation of Lol p 1 and Lol p 5 achieved 95.4% sensitivity and 96.4% specificity for ryegrass pollen sensitization.
Conclusion:
AeroDiff CytoBas has similar to superior diagnostic accuracy compared to singleplex IgE serology, with the additional advantage of a single assay to evaluate multiple allergens. This enables precise and efficient component-resolved diagnosis of aeroallergen sensitization to guide personalized treatment for patients with allergic rhinitis and/or asthma.
