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2021 OPEN ACCESS

Celakovska; Atopic Dermatitis and Sensitisation to Molecular Components of Alternaria, Cladosporium, Penicillium, Aspergillus, and Malassezia-Results of Allergy Explorer ALEX 2

Molecular diagnostic tests improve laboratory diagnostics of IgE-mediated allergies. Here, ALEX2- Allergy Explorer was used to determine the sensitisation profile towards moulds and yeast in patients suffering from atopic dermatitis. The relation between the sensitization to molecular components of moulds and yeast and the severity of atopic dermatitis, and the occurrence of bronchial asthma and allergic rhinitis was evaluated. Their results demonstrated that the sensitization to Mala s 6, Mala s 11, Sac c, Asp f 6, Cla h and Cla h 8 correlates to the severity of atopic dermatitis. The sensitization to Sac c, Alt a 6, Cla h, Cla h 8 was observed significantly more frequently in patients suffering from bronchial asthma to Mala s 6 in patients suffering from allergic rhinitis. In patients with severe form of atopic dermatitis (AD), a very high level of specific IgE was recorded to Mala s 11 (in 36%) and to Asp f 6 (in 12%)

doi: 10.3390/jof7030183. PMID: 33806376

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2021 OPEN ACCESS

Villalta; Evaluation and predictive value of IgE responses toward a comprehensive panel of house dust mite allergens using a new multiplex assay: a real-life experience on an Italian population

Specific IgE reactivity to a comprehensive panel of house dust mite allergens such as Der f 1, Der p 1, Der f 2, Der p 2, Der p 5, Der p 7, Der p 10, Der p 11, Der p 20, Der p 21 and Der p 23 was tested in patients with respiratory symptoms using Allergy Explorer (ALEX2®). Prevalence and IgE levels of Der f 1, Der f 2, Der p 1 and Der p 20 were significantly higher in asthmatic patients, whereas subjects negative for minor allergens resulted more frequently suffering from rhinitis. Asthmatic patients had IgE reactivity to a larger number of HDM allergens than patients with only rhinitis.

doi: 10.23822/EurAnnACI.1764-1489.195

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2021 NOT OPEN

Hoang; Modeling the conversion between specific IgE test platforms for nut allergens in children and adolescents

Plasma from Canadian and Austrian children/adolescents with peanut/tree nut sensitisation and a cohort of sensitised, high‐risk, pre‐school asthmatics were measured with three multiplex IgE test platforms: ALEX®, MeDALL‐chip (Mechanisms of Development of Allergy) (Thermo Fisher), and EUROLINE (EUROIMMUN). Using two regression methods, they demonstrated the ability to model allergen‐specific relationships for peanut and tree nut sIgE testing at the extract and molecular‐level, in order from highest to lowest: Ara h 2, Ara h 6, Jug r 1, Ana o 3, Ara h 1, Jug r 2, and Cor a 9.

doi: 10.1111/all.14529

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2021 OPEN ACCESS

Pinheiro; En route to personalized medicine: uncovering distinct IgE reactivity pattern to house dust mite components in Brazilian and Austrian allergic patients

A new in vitro multiplex allergen assay (ALEX) which allows simultaneous measurement of specific IgE against the whole allergen extract as well as its components were used. The data showed that in Brazil the inclusion of the molecular components Blo t 5 and/or Blo t 21 major allergens and Blo t 2 can increase the sensitivity and specificity of the assay for the diagnosis of allergy to B. tropicalis. Furthermore, monosensitisation to Blo t 2 was found in some individuals. Regarding the sensitization profile of individuals sensitized to D. pteronyssinus, they demonstrated that molecular components Der p23 and Der p 7, in addition to Der p 1 and Der p 2 are essential for an accurate diagnosis.

doi.org/10.1002/clt2.12004

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2020 NOT OPEN

Gonzáles-Pérez; Mite molecular profile in the Th2-polarized moderate-to-severe persistent asthma endotype subjected to high allergen exposure

González-Pérez et al. confirmed the dominant role of sIgE sensitization to Der p 2 and Der p 23 (86%) and Der p 1 (74%) among an asthmatic population in Spain with moderate-to-severe symptoms. Their results also demonstrated that more than 70% of the patients were sensitized to any of the major allergens, however an inclusion of allergens such as Der p 5, Der p 7 and Der p 21 enhanced diagnosis of mite-sensitized asthmatics up to >94%.

doi: 10.1159/000510118

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2020 OPEN ACCESS

Koch; IgE multiplex testing in house dust mite allergy is utile, and sensitivity is comparable to extract‐based singleplex testing

Another study compared allergy tests such as singleplex assay ImmunoCAP® (detecting Der p 1, 2, 10 and 23), the multiplex assay ImmunoCAP® ISAC (detecting Der p 1, 2 and 10) and Allergy Explorer (ALEX®) versions 1 and 2 (detecting Der p 1, 2, 5, 7, 10, 11, 20, 21 and 23). In general, sensitivity of molecular tests was lower compared to singleplex extract-based testing. However, sensitivity increased the more HDM allergen molecules were included in the test. Therefore, sensitivity of ALEX2® was higher compared to all other molecular assays. A clear correlation according to sensitivity between molecular test systems with HDM sIgE levels was observed implicating that higher sIgE levels lead to better sensitivity of molecular testing. Der p 1, 2 and 23 were confirmed to be major allergens. Moreover, Der p 23 is the third major HDM allergen and therefore essential to include in molecular test systems. Importantly, ALEX2® performed statistically equal to extract-based diagnosis in patients with allergen-specific IgE levels higher that 1.0 kU/L with a sensitivity of 97.2%.

doi: 10.1111/all.14271

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2020 OPEN ACCESS

Gellrich; De novo sensitization during subcutaneous allergen specifc immunotherapy - an analysis of 51 cases of SCIT and 33 symptomatically treated controls

To study the risk of subcutaneous allergen specific immunotherapy (SCIT)-induced neosensitisations, total IgE, sIgE reactivity and sIgG patterns to purifed natural allergen extract and allergen components were retrospectively measured using ALEX®. In 24% of SCIT-treated patients, they found new sIgE against allergen components of the allergen source treated by SCIT. In the SCIT-treated group, neosensitisation affected major and minor allergen components, and was accompanied by a strong induction of sIgG against major components. Patients undergoing SCIT might carry an enhanced risk of neosensitisation towards formerly unrecognized allergen components. According to anamnestic data, these neosensitisations might be of clinical relevance - supporting attempts towards personalized recombinant vaccines.

doi: 10.1038/s41598-020-63087-4

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2020 OPEN ACCESS

Rauber; The honey bee venom allergen Api m 10 displays one major IgE epitope, Api m 10160‐174

Nowadays, honey bee venom (HBV) allergy is treated by allergen immunotherapy (AIT) whose efficacy is low. Api m 10 was identified as a major allergen in HBV allergy, however the quantity of Api m 10 in the venom is of low abundance (approx. 1%). This may explain a potential link between underrepresentation in several therapeutic HBV preparations and treatment failure in dominantly Api m 10 sensitised HBV-allergic patients. In theory, this lack couled be compensated by spiking therapeutic HBV preparations with rApi m 10. However, Api m 10 is easily degraded. Therefore, alternative strategies need to be developed. To characterize the IgE response to Api m 10 in detail, linear IgE epitopes recognized by patients allergic to HBV were investigated. Synthetic peptides spanning the whole amino acid sequence of Api m 10 were coupled to macroarrays (Macro Array Diagnostics). Interestingly, one peptide, P54 (Api m 10160‐174; amino acid sequence: ADSDVTTLPTLIGKN), was recognized by 100% of the Api m 10‐positive sera and hence represents the dominant linear IgE epitope of Api m 10. A short immunodominant peptide is easier to produce, can be included in therapeutic preparations and thus may have relevant advantages over the use of full‐length rApi m 10.

doi: 10.1111/all.14187

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2020 OPEN ACCESS

Kowalczyk; Anaphylaxis in an 8-Year-Old Boy Following the Consumption of Poppy Seed

This case study describes a young patient with poppy seed allergy. Hazelnut allergy was found to co-occur in this patient. However, the primary responsible allergen was considered to be poppy seed.

doi: 10.18176/jiaci.0493

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2019 NOT OPEN

Scala; β‐1,3‐glucanase rOle e 9 and MnSOD rAsp f 6 IgE reactivity are the signature of atopic dermatitis in the Mediterranean area

To identify patterns of IgE sensitisation in patients with atopic dermatitis, ImmunoCAP ISAC® and Allergy Explorer‐ALEX® microarray analysis were performed. The olive tree pollen β‐1,3‐glucanase rOle e 9 and the manganese superoxide dismutase from Aspergillus rAsp f 6 were the molecules most significantly associated with AD occurrence and allowed to discriminate among the moderate and severe forms of disease. This publication demonstrates the importance of allergy tests using a large panel of molecular components to unveils important associations between IgE reactivity profiles and AD clinical presentation.

doi: 10.1111/cea.13555

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2019 NOT OPEN

Buzzulini; Evaluation of a new multiplex assay for allergy diagnosis

In a selected population of 105 allergic patients, skin prick test (SPT), singleplex (ImmunoCAP®) and multiplex assays (Allergy Explerer ALEX®) were compared. IgE antibodies to pollen, dust mites and food allergens were detected. Their results demonstrated a substantial agreement between ALEX® and SPT. When IgE to extracts was detected, ALEX® and ImmunoCAP® showed comparable results for inhalant and food allergens. In addition, using molecular components a higher agreement was observed for inhalant and food allergen molecules. Finally, the authors appreciated that ALEX® comprises all advantages of a multiplex test, such as an easy to learn method and a precise identification of the disease-eliciting molecules. To their opinion, ALEX® gives a boost to research in Molecular Allergy.

doi: 10.1016/j.cca.2019.02.025

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2019 NOT OPEN

Hoang; Extract and component-specific sensitization patterns in Canadian moderate-to-severe preschool asthmatics

In a cohort of North American preschool asthmatics, peanut and animal allergens were identified as major allergen sources. The dataset of this study provides detailed information on molecular diagnosis in preschool asthma. In particular, seed storage proteins (Ara h 1, 2, 3, 6), uteroglobin from cat (Fel d 1), and lipocalin from dog (Can f 1) demonstrated the strongest linkage to clinical markers of asthma severity. Allergen molecules can be biomarkers for stratification and endotyping in future prospective trials.

doi: 10.1111/all.13927

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2019 OPEN ACCESS

Bojcukova; Comparison of two multiplex arrays in the diagnostics of allergy

Based on Heffler et al., the present study went a step further and performed a comparison between the results of ALEX® and ImmunoCAP® ISAC for specific allergen components (most common allergens in these patients) such as: IgE against tree pollen Bet v 1, grass pollen Phl p 1 and 5, house dust mite Der p 1 and 2 and cat Fel d 1. They concluded a good correlation between ALEX® and ISAC.

doi.org/10.1186/s13601-019-0270-y

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2018 OPEN ACCESS

Heffler; Extended IgE profile based on an allergen macroarray: a novel tool for precision medicine in allergy diagnosis;

A very interesting study of 43 patients with allergies comparing two multiplex test systems (ImmunoCAP® ISAC and ALEX®) based on allergen extracts and molecules, found a good correlation between these methods. Several advantages of ALEX® were monitored, such as the ability of measuring total IgE, the high number of 282 reagents (157 extractive allergens and 125 molecular components) compared to 112 different components (both extracts and molecular allergens) in ISAC as well as the inhibition of CCD (Cross-reactive Carbohydrate Determinants) reactivity further improves the specificity of the IgE assay.

doi: 10.1186/s40413-018-0186-3

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