BACKGROUND: The identification of the primary sensitizing pollen is difficult in Southern European patients with Seasonal Allergic Rhinitis (SAR) if sensitized to various pollen sources with overlapping seasonality. A more precise diagnosis is obtained by IgE assays to allergen molecules, currently available as singleplex or microarrays. OBJECTIVES:To test the analytical performance of a multi-parameter immunoblot molecular "Pollen Test" specifically designed to test IgE antibodies to pollen extracts and molecules clinically relevant in Southern Europe. METHODS: Sera were obtained from 101 children and 98 adults with SAR and tested with a customized multiplex immunoblot assay (EUROLINE Southern European Pollen Profile [ESEP]; EUROIMMUN AG, Luebeck, Germany) containing a comprehensive panel of allergen extracts and molecules. ESEP's outcomes were then compared in selected sera (ESEP positive to negative = 2:1) with those of singleplex IgE assays (ImmunoCAP; ThermoFisher Scientific, Uppsala, Sweden). For each of the examined reagents, qualitative (sensitivity, specificity, accuracy), semi-quantitative (classes) and quantitative (Spearman's rank correlation, Bland-Altmann plots) comparisons were performed. RESULTS: Compared to ImmunoCAP, cumulative ESEP's sensitivity and specificity were 87% (95% CI 84%-90%) and 88% (83%-93%) for extracts and 99% (98%-100%) and 87% (83%-91%) for molecules. Cohen's kappa coefficients (κC ) ranged for extracts from 0.18 (Pellitory) to 0.50 (Cypress) and for molecules from 0.21 (Ole e 1) to 0.68 (Phl p 7). The quantitative outcomes of the two diagnostic tests were highly correlated, with Spearman's rank correlation coefficients always exceeding 0.80. Bland-Altmann plots showed a tendency of ESEP to overestimate serum specific IgE levels, when compared to ImmunoCAP. CONCLUSIONS AND CLINICAL RELEVANCE: Sensitivity and specificity of ESEP in testing serum IgE antibodies against pollen allergen extracts and molecules, in Italian patients with SAR, both exceeded 85%. The advantages and limitations of a multiplex customized immunoblot assay, in the routine clinical use of molecular diagnostics in Southern European pollen allergic patients, deserve to be tested.

A new molecular multiplex IgE assay for the diagnosis of pollen allergy in Mediterranean countries: a validation study / Di Fraia, Marco; Arasi, Stefania; Castelli, Sveva; Hofmaier, Stephanie; Potapova, Ekaterina; Villalta, Danilo; Tripodi, Salvatore; Sfika, Ifigenia; Zicari, Anna Maria; Villella, Valeria; Perna, Serena; Travaglini, Alessandro; Verardo, Pier Luigi; Matricardi, Paolo Maria. - In: CLINICAL AND EXPERIMENTAL ALLERGY. - ISSN 0954-7894. - ELETTRONICO. - (2018), pp. 1-9. [10.1111/cea.13264]

A new molecular multiplex IgE assay for the diagnosis of pollen allergy in Mediterranean countries: a validation study

Di Fraia, Marco;Zicari, Anna Maria;Villella, Valeria;Perna, Serena;Travaglini, Alessandro;Matricardi, Paolo Maria
2018

Abstract

BACKGROUND: The identification of the primary sensitizing pollen is difficult in Southern European patients with Seasonal Allergic Rhinitis (SAR) if sensitized to various pollen sources with overlapping seasonality. A more precise diagnosis is obtained by IgE assays to allergen molecules, currently available as singleplex or microarrays. OBJECTIVES:To test the analytical performance of a multi-parameter immunoblot molecular "Pollen Test" specifically designed to test IgE antibodies to pollen extracts and molecules clinically relevant in Southern Europe. METHODS: Sera were obtained from 101 children and 98 adults with SAR and tested with a customized multiplex immunoblot assay (EUROLINE Southern European Pollen Profile [ESEP]; EUROIMMUN AG, Luebeck, Germany) containing a comprehensive panel of allergen extracts and molecules. ESEP's outcomes were then compared in selected sera (ESEP positive to negative = 2:1) with those of singleplex IgE assays (ImmunoCAP; ThermoFisher Scientific, Uppsala, Sweden). For each of the examined reagents, qualitative (sensitivity, specificity, accuracy), semi-quantitative (classes) and quantitative (Spearman's rank correlation, Bland-Altmann plots) comparisons were performed. RESULTS: Compared to ImmunoCAP, cumulative ESEP's sensitivity and specificity were 87% (95% CI 84%-90%) and 88% (83%-93%) for extracts and 99% (98%-100%) and 87% (83%-91%) for molecules. Cohen's kappa coefficients (κC ) ranged for extracts from 0.18 (Pellitory) to 0.50 (Cypress) and for molecules from 0.21 (Ole e 1) to 0.68 (Phl p 7). The quantitative outcomes of the two diagnostic tests were highly correlated, with Spearman's rank correlation coefficients always exceeding 0.80. Bland-Altmann plots showed a tendency of ESEP to overestimate serum specific IgE levels, when compared to ImmunoCAP. CONCLUSIONS AND CLINICAL RELEVANCE: Sensitivity and specificity of ESEP in testing serum IgE antibodies against pollen allergen extracts and molecules, in Italian patients with SAR, both exceeded 85%. The advantages and limitations of a multiplex customized immunoblot assay, in the routine clinical use of molecular diagnostics in Southern European pollen allergic patients, deserve to be tested.
2018
allergen-specific immunotherapy; component-resolved diagnostics; immunoglobulin E; pollen; precision medicine; seasonal allergic rhinitis
01 Pubblicazione su rivista::01a Articolo in rivista
A new molecular multiplex IgE assay for the diagnosis of pollen allergy in Mediterranean countries: a validation study / Di Fraia, Marco; Arasi, Stefania; Castelli, Sveva; Hofmaier, Stephanie; Potapova, Ekaterina; Villalta, Danilo; Tripodi, Salvatore; Sfika, Ifigenia; Zicari, Anna Maria; Villella, Valeria; Perna, Serena; Travaglini, Alessandro; Verardo, Pier Luigi; Matricardi, Paolo Maria. - In: CLINICAL AND EXPERIMENTAL ALLERGY. - ISSN 0954-7894. - ELETTRONICO. - (2018), pp. 1-9. [10.1111/cea.13264]
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Utilizza questo identificativo per citare o creare un link a questo documento: https://hdl.handle.net/11573/1138158
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