Publications

The present study is the first investigating that the ALEX2 Allergy Xplorer is a reliable and useful stand-alone tool in the bottom-up for the diagnosis of tree nut allergy in children. In 169 children, skin prick tests, extract- and molecular allergen-specific IgE antibodies were examined for clinical relevance.

In singleplex tests, rAna o 3, nCor a 9/rCor a 14, and nJug r 1/nJug r 4 were found to be important in predicting clinical reactivity to cashew/pistachio, hazelnut, and walnut, respectively. However, disadvantages of singleplex tests include high costs and excessive blood sampling in multi-sensitised patients, and a limited number of components.

Using the ALEX2 Allergy Xplorer, tree nut allergens were found to be predictors for clinical reactivity. In detail, rPis v 1/rAna o 3, rPis v 1/nPis v 2/nPis v 3/rAna o 3, nCor a 9/nCor a 11/rCor a 14, and nJug r 1/nJug r 2/nJug r 4/nJug r 6 were significant in predicting clinical reactivity to cashew, pistachio, hazelnut, and walnut, respectively. However, there was moderate variability in the cut-off levels of these components. It seems logical to use a few empirical cut-off levels for different TN components. Hence, the investigation of different cut-off values and combinations of the components with SPT, sIgE and each other revealed that at the threshold of 1.0 kAU/L, rPis v 1 and rCor a 14 optimised predicting clinical reactivity to pistachio and hazelnut with 85.3 and 89.3% accuracy, respectively. A combination of nJug r 1 and nJug r 2 positivities had highest accuracy (89.5%) in predicting clinical reactivity to walnut. Furthermore, all patients with higher concentrations of rPis v 1 (≥15.0 kUA/L), rCor a 14 (≥5 kUA/L) and nJug r 1/nJug r 2 (≥15 kUA/L) were clinically reactive to pistachio, hazelnut and walnut, respectively.

doi: 10.1016/j.alit.2021.10.001

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Aytekin ES et al; Clin Exp Allergy 53(10):1041-1044, 2023; doi: 10.1111/cea.14350

Background

Molecular allergy diagnostics, such as the ALEX² test, offer a more refined approach for allergy diagnostics by reducing the need for oral food challenges and enabling the simultaneous detection of IgE reactivity to a broad panel of allergens. This study evaluated the diagnostic accuracy of ALEX² compared to conventional serological and skin tests. Therefore, a total of 123 Turkish children with peanut sensitisation were assessed.

Key findings:

31 (25%) children were clinically reactive to peanut, while 92 (75%) were tolerant and subsequently grouped
Ara h 2 and Ara h 6 were detected in 83.9% and 77.4% of the clinically reactive group, but only in 15.2% and 10.9% of the tolerant group
For Ara h 1 and Ara h 3, detection rates were 96.8% and 90.3% in the reactive group, compared to 70.7% and 37.0% in the tolerant group
Considering 2S albumin sensitisation, IgE reactivity was 9.7% to Ara h 2 only, 3.2% to Ara h 4 only, 74.2% to both, and 12.9% to neither in the clinically reactive group
The 12.9% not sensitised to Ara h 2 & 6, were also not sensitised to Ara h 1 & 3
Ara h 8 was detected in 3 peanut-tolerant patients, while Ara h 9 was detected in 48.4% of the clinically reactive group & 38.0% of the tolerant group
Comparison of AUCs showed similar diagnostic potential for peanut allergy, with no significant differences between SPT (AUC = 0.927), peanut extract sIgE (AUC = 0.854), and IgE to Ara h 1, 2, 3, and 6 (AUCs = 0.829–0.882)
Combining diagnostic tests improved peanut sensitisation accuracy
The combination of Ara h 2, Ara h 6 and SPT had the best accuracy (90.9%) with the highest likelihood ration (LR+; 22.2)
While Ara h 2 and Ara h 6 showed the best accuracy for predicting clinical reactivity, peanut-allergic patients had higher IgE reactivity to Ara h 1 and Ara h 3

Conclusion:

This study confirms that IgE antibodies to Ara h 1, 2, 3, and 6 predict peanut allergy in children. The combination of SPT with sIgE to Ara h 2 and Ara h 6 achieved over 90% diagnostic accuracy, thus showing that the ALEX² test complements the SPT.

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Biliute G et al; Clin Transl Allergy. 14(1):e12332, 2024; doi: 10.1002/clt2.12332

Background

House dust mites (HDMs) are major triggers for respiratory allergic diseases and dermatitis. Given the global prevalence of HDM allergy and its varying sensitisation profiles, this study aimed to assess the molecular allergen sensitisation patterns in the Lithuanian population, particularly focusing on Dermatophagoides pteronyssinus (Der p), and to explore patterns of concomitant reactivity among various allergens to improve HDM allergy diagnostics. Sensitisation patterns to major (Der p 1, Der p 2, and Der p 23) and minor (Der p 5, Der p 7, and Der p 21) Der p allergen components, along with allergen components from other sources, including tropomyosins (Der p 10, Per a 7, Pen m 1, Ani s 3, Blo t 10) and arginine kinases (Pen m 2, Bla g 9, Der p 20), were evaluated using the ALEX² test. A total of 1,520 individuals, both children and adults, with suspected atopic disease were examined.

Key findings:

1,060 (69.74%) patients showed sensitisation to inhalant, food, or insect allergens
Thereof, 481 individuals (45.38%) exhibited sensitisation to at least one Der p allergen component
Within the sensitised group, 37.21% were sensitised to Der p 5, 7, or 21 in addition to major allergenic components
Different age groups showed distinct sensitisation patterns: age-related influence
Cross‐reactivities were seen between Der p 5 & Blo t 5 and Der p 21 & Blo t 21
3.40% showed sensitisation to tropomyosins, and 1.79% showed sensitisation to Der p 10
Sensitisation to arginine kinase molecular allergens (Pen m 2, Bla g 9, Der p 20) was rare in the studied population, with only 4.53% (n = 48) of all sensitised patients demonstrating a reaction to arginine kinase

Conclusion:

This study highlights significant differences in sensitisation profiles to Der p allergens within the Lithuanian population, influenced by geographical, population-specific, and age-related factors. Understanding sensitisation patterns to major and minor HDM allergen components is crucial for improved diagnosis and management of HDM‐related allergic diseases. Additionally, the study demonstrates that understanding concomitant reactivity among allergens such as Der p 5 and Blo t 5, Der p 21 and Blo t 21, as well as tropomyosins and arginine kinases, is vital for enhancing diagnostic strategies and developing targeted interventions for individuals with allergies.

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Summary:

This research sheds light on the complex patterns of allergy sensitisation and their implications for medical needs, highlighting the importance of understanding molecular allergens and their role in allergies. It employed a European Health Examination Survey cohort (n=1462) of Luxembourg, deep personal sIgE profiling, and an unsupervised computational method.

Among the key findings:

42.6% of participants reported physician-diagnosed allergies, with 44% testing positive for IgE against at least one allergen or extract.
Sensitisation sources were primarily tree pollens (52.4%), grass pollens (51.8%), and mites (40.3%).
The youngest group of participants (25–34 years old) represented a significant cluster (24.4%) with multi-sensitisation to respiratory sources which reflects the highest medical needs. Poly-sensitisation, especially in younger individuals, was linked to more severe symptoms and potential increased socioeconomic costs.
The study found that participants with respiratory allergies often exhibited sensitisation to multiple molecular allergens within the same group, indicating a high likelihood of clinical reactivity, particularly evident in complex IgE patterns for grass pollen allergens like Phl p 1, 2, 5, and 6. This phenomenon of antibody responses evolving to complex patterns, known as "molecular spreading," was linked to grass pollen allergy, starting with the initiator allergen Phl p 1 and subsequently expanding to include Phl p 4, 5, and other allergens.
Certain initiator allergens, such as Phl p 1 and Bet v 1, played central roles in the progression of atopy (genetic predisposition to allergies).
The study's network analysis revealed a significant interconnectedness of the PR-10 allergen cluster with many other clusters. Notably, this central role was confirmed as the PR-10 allergens were found to coexist with other allergens from another cluster rather than forming a separate isolated cluster.
The largest cluster, including a quarter of sensitised participants, was characterised not only by allergens from the PR-10 protein family but also by sensitisation to airborne signifier allergens from other clusters, such as grass pollen allergens (Phl p 1, Lol p1, Phl p 5) and house dust mite allergens (Der p 2, Der f 2, Der p 1).
The study's strengths included a deep population-based knowledge database and an extensive IgE panel for analysing complex IgE profiles.
The study utilised the ALEX² Allergy Xplorer, which provided an extra 200 data points compared to the Immuno Solid-phase Allergen Chip. These additional data points were instrumental in defining IgE clusters, including specific allergens like Fag s 1, Cor a 1.0103, Fra a 1/3, Der f 2, Der p 23, Der p 7, and Gly d 2.
Importantly, the study's molecule-resolved approach ensured comparability with other research studies, enhancing the depth and precision of their analysis.

https://doi.org/10.1002/clt2.12292

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Eidukaite A et al; World Allergy Organ J. 4;16(10):100827, 2023; doi: 10.1016/j.waojou.2023.100827

Background

Pet allergens are among the most common inhalant allergens causing allergic diseases. In recent years, there has been an increasing trend in asthma and allergic rhinitis cases due to allergy to pets. This study determined the molecular sensitisation profile to cat and dog allergens in Lithuanian children with allergy-like symptoms. A total of 574 children (0-18 years) with suspected atopic conditions were tested with ALEX². The 433 children sensitised to at least one allergen were further analysed for sensitisation to cat (Fel d 1, Fel d 2, Fel d 4, and Fel d 7) and dog (Can f 1, Can f 2, Can f 3, Can f 4, Can f 5, and Can f 6) allergen components.

Key findings:

A high sensitisation frequency to different inhalant allergens in Lithuanian children was observed: pollen (66.3%), dust mites (44.1%), and molds (24.3%)
More than half of the children (n=247, 57.04%) tested positive to at least one dog or cat allergen component
9.71% were sensitised to dog components, 29.14% to cat components and 61.13% to components from both sources
The major sensitisers were Fel d 1 (84.8%) and Can f 1 (59.4%)
5.7% patients were sensitised both to Fel d 1 + Fel d 4 and Can f 1/2 + Can f 5, indicating the high risk of severe asthma
Monosensitisation to Fel d 1 was the dominant pattern among the children (26.3%)
42.9% patients were sensitised to 3 or more different mammalian protein families and 40.4% to 3 or more lipocalins

Conclusion:

This study reveals a high rate of sensitisation to inhalant allergens among Lithuanian children, with dog and/or cat allergens being particularly prevalent. Most children were polysensitised to both cat and dog allergens, though sensitisation to cat allergens was more common. In conclusion, in vitro molecular allergy diagnostics is a useful tool for accurate true sensitisation diagnosis and prognosis of disease severity.

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Hamilton RG et al; J Asthma 1-8, 2024; doi: 10.1080/02770903.2024.2324839

Background:

Identifying a patient’s atopic status is a key factor in the management of asthma. The qualitative Phadiatop, a multi-aeroallergen singleplex screening assay by ImmunoCAP, has been used to differentiate atopic from non-atopic asthma since 2010. The quantitative multiplex array ALEX² is emerging as a promising alternative, potentially addressing limitations such as the restricted profile of 10 aeroallergen specificities found in Phadiatop. This study aimed to evaluate the strengths and weaknesses of ALEX² in screening of asthma patients for atopic status. For this purpose, the atopic status of 42 asthmatic Amish and Hutterite children with equivocal and positive Phadiatop results was assessed by ALEX².

Key findings:

In total, 42 asthmatic children were analysed by Phadiatop and total IgE was measured
22 children with negative Phadiatop (<0.1 kUA/L) and total IgE <100 kU/L were defined non-atopic and thus, excluded from ALEX² testing
Among 3 children with a negative Phadiatop but total IgE >100 kU/L and 6 children with equivocal Phadiatop (0.1-0-2 kUA/L), 44% had detectable IgE by ALEX² to mite, tree pollen, and other allergens (not detected by Phadiatop)
Of 11 children with positive Phadiatop (>0.2 kUA/L), all but one were positive by ALEX²
IgEs against mold and rabbit aeroallergens corresponded well with the children’s agricultural and pet exposure history
3 children were sensitised to profilin, nsLTP, or PR-10 protein families

Conclusion:

ALEX² macroarray containing allergen extracts and molecules proved to be a good alternative to Phadiatop in the assessment of atopic status of asthma. Screening of asthmatics for atopy with ALEX² assay instead of Phadiatop enhanced the knowledge about the IgE asthma triggers as well as genuine versus cross-reactive allergen specificities. The combination of ALEX² testing, traditional single-plex assay and patient’s clinical history provides a better interpretation of sensitisation patterns and planning of treatment strategies.

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Kalyniuk V et al; Frontiers in Allergy Vol. 5, 2024; doi: 10.3389/falgy.2024.1438393

Background

Sensitisation to fungi is a significant factor in allergic rhinitis and asthma. Moreover, it has been shown that a large proportion of severe asthmatics are sensitised to fungal allergens such as Alternaria and Aspergillus. This study investigated the sensitisation profiles of patients hypersensitive to fungal allergens in the Ukrainian population, examine their co-sensitisation to other allergens, and explore the relationships between different allergens in provoking hypersensitivity. Children and adults (n= 3,349; 77.84% children vs. 22.16% adults) sensitive to fungal components were tested via ALEX² in 17 regions of Ukraine. Eligible participants had a history of allergic rhinitis, chronic urticaria, allergic dermatitis, or asthma along with sensitisation to fungal allergens.

Key findings:

79.30% were sensitive to Alt a 1 and 62.09 % showed Alt a 1 mono-sensitisation
Fungi-polysensitised patients were found to have unique allergological profiles, with 84.77% reacting to two or more allergens from various groups
The immune response to Alt a 1 may trigger sensitisation to various other common allergens, increasing the likelihood of sensitivity to grasses (primarily to Phl p 2), ragweed (Amb a 1), pectate lyase (Cry j 1), and the cat allergen Fel d 1
Fungi-polysensitised individuals were often sensitised to cross-reactive molecules, like lipocalins (Fel d 4 and Can f 6)
Ambrosia extract (Amb a) played a significant role in ragweed pollen sensitisation, while Phl p 2 might be important in grass sensitivity in patients with Alternaria allergy

Conclusion:

The study highlighted the participants' unique allergenic profiles, which often included multiple allergens from various sources. These findings underscore the importance of considering fungal allergens within the broader scope of allergic diseases, particularly due to their potential to enhance responses to other non-fungal allergens. In conclusion, this study emphasizes the importance of in-vitro multiplex allergy test such ALEX, which allows measurements of specific IgE against a large number of allergen extracts and molecular allergens, in patients with sensitisation to fungi.

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Knyziak-Mędrzycka I et al; Int J Mol Sci. 25(2):825, 2024; doi: 10.3390/ijms25020825

Background

Food allergies are becoming more prevalent. Environmental factors play a significant role in allergy development, leading to diverse sensitisation patterns influenced by geographic regions, climate, dietary habits, and/or food preparation methods. The aim of this study was to conduct an in-depth evaluation of paediatric sensitisation to food allergens, with a specific emphasis on the eight major food allergens (the “Big 8”): cow milk, eggs, wheat, soybeans, fish, crustacean shellfish, tree nuts, and peanuts. To achieve this, the prevalence and serum levels of sIgE from 3,715 children and adolescents across Poland were analysed via ALEX, providing a comprehensive review of IgE sensitisation profiles based on both molecular and extract analyses of food allergens.

Key findings:

572 results were obtained with ALEX, and 3,143 results with ALEX2
The final analysis included the sIgE results obtained with 95 food extracts and 77 food allergen molecules
The highest rates of sIgE to food allergen extracts were to peanuts (29.20%), hazelnuts (28.20%), and apple (23.60%)
The highest rates of sIgE to allergenic molecules were to PR-10 family (Cor a 1.0401 (23.77%), Mal d 1 (22.37%), Ara h 8 (16.93%)), and globulin 7/8S (Ara h 1 (15.59%)
The lowest rates of sIgE to extracts were to strawberry (0.40%), oregano (0.30%), and thornback ray (0.16%)
The lowest rates of sIgE to allergenic molecules were to Mal d 2 (0.27%) (TLP), Ani s 1 (0.30%) (Kunitz-type serine protease inhibitor), and Che a 1 (0.43%) (Ole e 1 family)
The sensitisation rates to storage proteins decreased significantly with age, while the sensitisation rates to PR-10 family proteins increased significantly with age

Conclusion:

The multiplex ALEX® test showed unique molecular sensitisation profiles to major food allergens in Polish children, differing significantly from sensitisation profiles reported in other countries. Peanut, hazel, and apple consistently ranked among the top allergens, regardless of whether extracts or molecules were tested. Furthermore, serum sIgE analysis across all regions of Poland revealed an age-dependent variation in the molecular sensitisation profiles to food allergens.

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Background:

Allergic rhinoconjunctivitis affects 20% of the general population. The use of native allergen extracts decreases diagnostic specificity of pollen allergy due to CCDs (Cross-reactive carbohydrate determinants) and panallergens (profilins and polcalcins) which can hamper or confound test results. Thus, allergen extracts bear the significant risk of misdiagnosis. The aim of the present study is to determine the prevalence of multiple pollen sensitisations and cross-reactivity and to evaluate the sensitivity and specificity of molecular allergy diagnostics. This study investigated the prevalence of sensitisation to seasonal pollen extracts, cross-reactive carbohydrate determinants (CCDs), profilins, and polcalcins in a group of 2948 patients. The patients were screened for specific immunoglobulin E to ash, birch, mugwort, ragweed, and timothy grass pollen extracts. The patients were grouped based on the number of positive tests (ranging from 1 to 5), and a subset of 742 patients was used to determine the sensitivity and specificity of molecular allergy diagnosis (MAD) using commercially available test methods (ALEX2 Allergy Xplorer and ImmunoCAP).

Key findings:

Prevalence of Sensitisation:

Remarkably, the correlation of sIgE extract and skin prick test results demonstrated that sIgE positive/prick negative patients were frequently observed for pollen allergens. 13.3% of the positive extract determinations had negative corresponding prick test results.
55.2% of the patients were positive to at least one of the five pollen allergens investigated
34.0% of the patients were positive to multiple pollen allergens
18.5% of pollen-sensitised patients had sensitisation to CCDs or panallergens (profilins or polcalcins).

Specific Sensitisation Rates:

Sensitisation to CCDs was observed in 8.7% of pollen-sensitised patients
Sensitisation to profilins was observed in 10.9% of pollen-sensitised patients
Sensitisation to polcalcins was observed in 2.9% of pollen-sensitised patients

Molecular Allergy Diagnosis (MAD):

The study used MAD with specific allergen components (Fra/Ole e 1, Bet v 1, Phl p 1, Art v 1, and Amb a 1).
The sensitivity of MAD was high, ranging from 92% to 100% using ALEX2.
Specificity was 100%.

Conclusions:

Cross-reactivity, particularly with CCDs, profilins, and polcalcins, poses a risk of misdiagnosis in about one-fifth of pollen-sensitised patients. Despite cross-reactivity challenges, MAD using specific allergen components was found to be sensitive and specific. In summary, the study highlights the importance of considering molecular components in allergy diagnosis to enhance specificity and reduce the risk of misdiagnosis in patients with pollen allergies.

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Short summary: In this study, a mathematical model for predicting the probability of asthma control based on the FeNO dynamics after 3 months of treatment, the number of cat allergens to which sensitisation is detected and the duration of exposure to a domestic cat after molecular allergy testing was developed.

Summary: The aim of this study was to develop a model for predicting asthma control status in school-aged children, who are sensitised to cat allergens. In this retrospective study, 302 patients were included. Inclusion criteria involved diagnosis of bronchial asthma, age 6-17 years and sensitisation to at least one of the following cat allergens Fel d 1, Fel d 2, Fel d 4, Fel d 7. Patients were selected upon careful history taking, spirometry, skin-prick test with Fel d 1 standardised extract (5,000 BAU / mL), measurement of total IgE and fractional exhaled nitric oxide (FeNO) before and after 3 months of treatment.

To obtain a comprehensive sensitisation profile, serum samples were analysed using the ALEX² test. The following sensitisation prevalences were measured:

• Fel d 1 in 291 (96.36%) children

• Fel d 2 in 18 (5.96%) children

• Fel d 4 in 59 (19.54%) children

• Fel d 7 in 75 (24.83%) children

Combined sensitisations were also observed:

• Fel d 1, Fel d 7 in 32 (10.60%) patients

• Fel d 1, Fel d 4 in 22 (7.28%) patients

• Fel d 1, Fel d 4, Fel d 7 in 22 (7.28%) patients

• Fel d 1, Fel d 2, Fel d 4, Fel d 7 in 12 (3.97%) patients

To evaluate the correlation between controlled asthma and sensitisation to cat allergens, the Searman’s rank correlation coefficient was calculated. A correlation was found between the presence of asthma control and the number of cat allergens to which the child is sensitised, which emphasises the importance of multiple sensitisations. Regarding the structure of sensitisation, a correlation was found between the presence of asthma control and sensitisation to Fel d 4 and Fel d 7. According to the Consensus document on dog and cat allergy, sensitisation to two or more allergens is associated with more severe respiratory symptoms and is a marker of severe asthma.

doi: 10.36740/WLek202206110

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Short summary:

This study is the first to evaluate potential differences in IgE sensitisation profiles of cockroach-sensitised patients with perennial respiratory allergy symptoms from two different geographic areas: coastal China (Hong Kong, HK) and central Europe (Austria). Molecular profiling using the ALEX² Allergy Xplorer identified differences between cockroach-sensitised allergic patients from HK and Austria in terms of primary sensitisers and molecular IgE reactivity patterns. Tropomyosin from American cockroach (Per a 7) was shown to be significantly associated with asthma symptoms and might be suitable as a biomarker for more severe respiratory allergy symptoms.

Summary:

In addition to skin prick testing and/or IgE reactivity with cockroach extract, using the ELISA-based multiplex allergy test ALEX² comprising a broad panel of cockroach allergens, the aim was to assess the patients’ molecular IgE reactivity patterns and to determine the primary sensitising allergen sources and to differentiate between co- and cross-sensitisations in the studied cohorts. Molecular IgE reactivity profiles were analysed via multiplex assay for sensitisation to allergens and extracts from cockroach, house dust mite, shellfish, and 3 additional insect species. In the HK group, genuine sensitisation to cockroach was found in 45%, but none of the patients in the Austrian cohort was truly sensitised to that allergen source, thus cockroach is not a relevant primary sensitising allergen source in Austria. Most patients from HK were cross-sensitised to other insects and/or shellfish, presumably by broad reactivity to tropomyosin and arginine kinase. About half of the Austrian subjects lacked IgE to these pan-allergens, indicating co- but not cross-sensitisation to insects and/or shellfish. Regarding IgE recognition frequencies, arginine kinases (64% HK, 10% Austria) and tropomyosins (42% HK, 15% Austria) were most frequently recognised; Bla g 4 (lipocalin) was detected in HK patients only (42%). IgE sensitisation to Per a 7 was significantly higher in patients with asthma than patients without asthma. Sera from HDM-sensitised subjects from HK showed a higher proportion of sensitisation to minor mite allergens.

doi: 10.1016/j.jacig.2022.04.003

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Reznik Y. V. et al; Regul. Mech. Biosyst. 14(4), 2023; doi: 10.15421/022391

Background

Fungi are ubiquitous microorganisms found in both indoor and outdoor environments. They play significant roles in ecology and agriculture but also pose health risks, particularly through fungal allergies, which are among the most common health issues globally. The primary focus of this study was to assess the sensitivity to fungal allergens in individuals from the Vinnytsya region. A total of 87 individuals, aged 1 to 66, were analysed for IgE-mediated genuine sensitisation to fungal allergens, including Alternaria (Alt a 1, Alt a 6), Cladosporium (Cla h, Cla h 8), Aspergillus (Asp f 1, Asp f 3, Asp f 4, Asp f 6), Penicillium (Pen ch), Malassezia (Mala s 11, Mala s 5, Mala s 6), and Saccharomyces (Sac c), using the ALEX² test.

Key findings:

Sensitivity to fungal allergens was identified in 20 patients, representing 23.0% of those tested
The highest level of sensitivity was observed to Alternaria fungus (in 75% of tested patients): 14 (93.3%) patients were sensitive to Alt a 1, one of whom was also sensitive to Alt a 6; o One patient was sensitive only to Alt a 6
Sensitivity to Aspergillus and Malassezia was the second highest, with 6 patients (30%) of sensitised individuals, reacting to these two fungal allergens: One patient was sensitised to Asp f 4; Two patients were sensitised to Mala s 6; Patients sensitised to Aspergillus were also co-sensitised to Alternaria, while those sensitised to Malassezia were co-sensitised to Alternaria, Aspergillus, and Cladosporium
Two patients were sensitised to Cladosporium
One patient was sensitised to Penicillium but no other fungal allergen

Conclusion:

This study revealed that Alternaria was the leading cause of sensitisation in the tested population of the Vinnytsya region, followed by Aspergillus allergens, which are clinically known to trigger both allergic and infectious reactions. Although Cladosporium typically has the highest spore counts in the air, it is characterised by low allergenicity. Elevated levels of specific and total IgE can act as predictive markers for potential fungal allergies, highlighting their significance in diagnosis and prevention. In this regard, the ALEX² test is particularly valuable, as it enables simultaneous measurement of total and specific IgE for a wide range of fungal allergens, aiding in the distinction between genuine sensitisation to fungal allergens and cross-reactive allergens.

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Rodinkova V et al; World Allergy Organ J. 17(5):100908, 2024; doi: 10.1016/j.waojou.2024.100908

Background

Fungi are able to trigger respiratory tract allergic diseases, yet there is a lack of sufficient data regarding individual sensitisation to fungal allergens. This study focused on analysing the epidemiological patterns of age-related, individual, and regional sensitisation to fungal allergens within the Ukrainian population. A total of 20,033 individuals, both children and adults, with a history of allergic rhinitis, atopic dermatitis and/or asthma were tested with ALEX². A complex of programs in the Python language was developed and Bayesian network analysis was applied to determine the sensitivity combinations in individual patients to various fungal components.

Key findings:

16.71% individuals showed sensitivity to at least one fungal allergen
Sensitivity to Alt a 1, either alone or in combination with other allergens, was observed in 79.39%, making it the most common sensitisation
62.17% individuals showed mono-sensitisation to Alt a 1
Mono-sensitisation to Mala s 6 was the second most common, at 4.06%
3.28% showed a combined sensitivity to Alt a 1 – Asp f 3
Pen ch and Cla h extracts stimulated the production of the lowest median sIgE levels
The highest median sIgE levels were for Alt a 1, Mala s 11 and Asp f 6
Median sIgE levels increased in adults compared to children for all Aspergillus fumigatus components, as well as for Mala s 5 and Mala s 11, but decreased for the other allergens
The sensitisation rates to fungi in general and specifically to Alternaria were lower in the western parts of Ukraine, especially in the Carpathian region
Bayesian modeling showed a 92.42% probability that sensitisation to Alt a 1 occurs when sensitivity to Cla h 8, Mala s 11, Mala s 5, and Mala s 6 is absent
A lack of sensitivity to one allergen often coincided with others, suggesting independent sensitisation to different fungal allergens

Conclusion:

This study showed that Alt a 1 sensitivity was predominant, with lower rates in the western regions of Ukraine. The highest median sIgE levels were induced by Alt a 1, Mala s 11 and Asp f 6. Bayesian analysis indicates a strong likelihood that sensitisation to different fungal allergens occurs independently. The idea that sensitisation to one allergen may be protective against sensitisation to others requires further investigations.

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Rostaher A et al, Animals (Basel), 2023; doi: 10.3390/ani13193002

Background

Hymenoptera venom allergy in dogs can lead to severe, life-threatening anaphylactic reactions following stings from bees, wasps, or ants. Venom immunotherapy (VIT) is the only known treatment to prevent further systemic responses. This study aimed to evaluate the safety and efficacy of VIT in ten dogs with a history of anaphylaxis following repeated Hymenoptera stings, all of which had positive intradermal tests for bee and/or wasp venom and elevated allergen-specific IgE levels. Allergen-specific IgE levels to bee and wasp allergen extracts were measured using the Allercept test. Additionally, the PAX (Pet Allergy Xplorer) test was used to measure specific allergen components including nApi m 1, rApi m 10, rApi m 2, rApi m 3, rApi m 5, rVes v1and rVes v 5.

Key findings:

In both Allercept and PAX, bee-specific IgE levels were significantly higher than wasp-specific IgE levels
Most bee-allergic dogs showed sensitisation to Api m 1, 2, 3, and 10, with no dogs showing sensitisation to Api m 5
Nine dogs with a history of anaphylaxis to a single insect species were found to be sensitised to both bee and wasp allergens when all test modalities were considered
Allergens used in VIT, based on history and allergological work-up, included bee (8 dogs), wasp (1 dog), and both (1 dog)
No systemic adverse events occurred during the induction and maintenance phases
VIT efficacy was assessed by observing clinical reactions to re-stings: of the seven re-stung dogs, only one developed mild angioedema at the sting site, while the others remained asymptomatic

Conclusion:

This study confirmed that VIT is a safe and effective treatment for Hymenoptera-allergic dogs, demonstrating efficacy similar to that in humans. Furthermore, both skin testing and sIgE tests were reliable in identifying the underlying insect sensitisation in these dogs.

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Summary:

This is the first paper documenting the use of the Pet Allergy XplorerTM in dogs with hymenoptera-venom induced anaphylaxis: Hymenoptera allergens, such as bee and wasp stings, can trigger anaphylaxis in both dogs and humans. While venom-specific immunotherapy (VIT) is an effective treatment for preventing life-threatening reactions in humans, there's a lack of prospective clinical data on its efficacy in dogs. This uncontrolled prospective clinical trial involved 10 dogs with a history of anaphylaxis due to Hymenoptera stings. The sensitization to bee and wasp allergens was demonstrated by intradermal testing (IDT) and allergen-speciﬁc IgE serology (Allercept®, Pet Allergy XplorerTM). As reported by the owners, 8 out of 10 dogs had a history of allergic reactions to bee stings, one dog was stung by a wasp, and one dog had a history of bee and wasp stings. All 10 dogs tested positive in IgE Allercept® and 9 out of 10 dogs in the PAX® serology test. When correlating the data of both serological tests, a statistically signiﬁcant moderate positive correlation was found (r = 0.62, p = 0.028) for bee allergens. Eighty percent of the dogs (8/10 dogs) tested positive for CCD IgE, as determined by Allercept® test, only two dogs were negative for these speciﬁc antibodies. The levels of bee- speciﬁc IgEs were signiﬁcantly higher than the wasp-speciﬁc IgE for Allercept® (median 350 vs. 30 HERBU/mL, p = 0.007) and PAX test (median 182 vs. 27 ng/mL, p = 0.003). Most of the bee-allergic dogs were sensitized to the following allergen components: Api m 1, Api m 2, Api m 3, and Api m 10. None of the dogs were sensitized to Api m 5. As there were only two dogs allergic to wasps, the dataset for the allergen components was too small for statistical analysis. VIT was administered in an induction phase followed by monthly maintenance injections, and it proved to be safe with no systemic adverse events. Among the re-stung dogs, only one developed mild angioedema, while the rest remained asymptomatic. These findings demonstrate that VIT is a safe and effective treatment for Hymenoptera-allergic dogs, potentially preventing anaphylactic reactions.

https://pubmed.ncbi.nlm.nih.gov/37835609/

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Scala E et al; Eur Ann Allergy Clin Immunol, 2023; doi: 10.23822/EurAnnACI.1764-1489.314

Background

Non-specific lipid transfer proteins (nsLTPs), which are important food and environmental allergens, are especially prevalent in areas with high plant food intake. Initially believed to be confined to the Mediterranean area, LTP IgE reactivity is now known to be widespread, with variations in epitope recognition among different populations, indicating the potential impact of individual, environmental, and dietary factors. This study aimed to investigate the individual LTP molecular recognition profiles, clinical outcomes, and potential influencing external factors in an Italian population with IgE reactivity to at least one of 15 LTPs. A total of 1,831 patients were screened using ALEX², and the 426 patients who exhibited at least one LTP reaction were included in the study.

Key findings:

Most common sensitisations were to: Pru p 3 (77%), Mal d 3 (60%), Zea m 14 (60%), Ara h 9 (50%), Pla a 3 (50%)
In the studied population, Pru p 3 remains the molecule most commonly associated with LTP allergy however, the results showed that Pru p 3 is less effective in detecting patients at high risk for severe reactions than specific sensitisations to Cor a 8, Mal d 3, or Arah 9
234 patients (54.9%) were solely sensitised to LTPs, while the remaining individuals were also sensitised to other pan-allergens: 118 (27.7%) to PR10, 56 (13.1%) to profilin, and 23 (5.4%) to polcalcin
Higher sIgE levels and concurrent sensitisation to >5 allergen molecules were significantly associated with an increased risk of severe reactions
Influence of co-factors on clinical outcomes:
Co-factors associated with an increased risk of severe reactions: mono-reactivity to LTP (44.6%; p=0.001), FDEIA (10.8%; p=0.001), and FDNIH (11.5%; p=0.005)
Co-factors associated with decreased symptom severity: reactivity to PR10 (24.2%; p=0.001), profilin hypersensitivity (10.3%; p=0.001), and/or atopic dermatitis (16.7%; p=0.001)

Conclusion:

In conclusion, LTP syndrome severity is linked to a broader IgE repertoire and higher IgE levels. Ara h 9, Cor a 8, and Mal d 3 were strongly associated with more severe symptoms. Further, co-factors such as FDEIA, FDHIH, and LTP monoreactivity can worsen patient clinical outcomes, while atopic dermatitis and co-sensitisation to profilin and/or PR10 can improve them. Thus, these co-factors can aid in the daily clinical management of LTP syndrome.

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Scala E et al; J Investig Allergol Clin Immunol Vol. 35(3), 2024; doi: 10.18176/jiaci.0986

Background

Edible insects are considered the next super global food as it provides a rich source of protein and micronutrients, despite the risk of anaphylaxis. In this study, the IgE-mediated sensitisation status of three insects (cricket, locust, and mealworm) in patients never knowingly consumed edible insects before was assessed. A total of 2,014 participants were screened with ALEX² for the extracts Acheta Domesticus (Ad), Locusta migratoria (Lm), and Tenebrio molitor (Tm).

Key findings:

9.7% participants were sensitised to insects, but none of these had knowingly consumed edible insects in the past
46% showed reactivity to all three extracts, and the mono-sensitisation rates were 18% for Ad, 15% for Tm, and 3% for Lm
40-60% showed sensitivity to allergenic components found in crustaceans, molluscs, and nematodes: Tropomyosin was co-recognised by 34%, and arginine kinase by 18.5%; <5% showed reactivity to Sarcoplasmic-CB, Troponin-C, Paramyosin, or Myosin-light-chain; 55.4% did not show any reactivity to invertebrate pan-allergens
16.9% experienced mono-sensitisation exclusively to insects
Arachnid reactivity was inversely proportional to insect allergen sensitisation, while Mollusca, Blattoidea, and tropomyosin reactivity displayed a direct relationship
Myosin-light-chain reactivity correlated with Ad and Lm, and Troponin-C with Ad and Tm sensitisation

Conclusion:

Many individuals with IgE sensitisation to edible insects had no prior exposure to them, leaving the cause of sensitisation unclear. This study revealed a direct link between insect sensitisation and reactivity to tropomyosin, mollusks, and cockroaches, suggesting these as primary sensitisers. With the growing acceptance and popularity of edible insects as a novel food source, it is crucial to address their potential allergenicity and related health concerns.

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Summary:

In the present study, in-vitro IgE tests using molecular components represent important innovations in the diagnosis of rare diseases, such as inborn errors with HIE (Hyper-IgE). Thus, IgE sensitisation profiles can be evaluated, even in cases in which the patient’s cognitive defect does not allow for correct allergological classification. The use of allergen multiplex tests improves both, the management of skin pathology and the quality of life of the individual patients.

Here, a proteomic test combined with IgE testing were performed to study inborn errors with an atopic dermatitis-like clinical picture associated with a deregulated IgE response. Four patients with rare diseases, such as recessive X–linked ichthyosis, Comel–Netherton syndrome, monosomy 1p36 syndrome, and a microduplication of Xp11.4 associated with extremely high levels of IgE were evaluated by comparative genomic hybridisation microarray analysis and specific IgE evaluation using allergen macroarray (ALEX2) and allergen microarray (ISAC).

In patients suffering from atopic dermatitis, IgE reactivity to LTP-related food allergens was detected, impacting the quality of life. The avoidance of LTP-related foods from the diet resulted in a marked improvement of the patient’s clinical condition.

Furthermore, reactivity to CCDs (Cross-reactive carbohydrate determinants) was evidenced in one patient. Although sensitisation to CCDs has no clinical impact, it is important to remember that IgE testing without blocking CCD-specific IgE antibodies can give false IgE test results.

In another patient, reactivity to autoreactive allergenic components was shown. Interestingly, the patient was positive for both Asp f 6 and Ole e 9 considered to be a signature of patients with atopic dermatitis in the Mediterranean area.

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Storz L et al; Front Med (Lausanne) 11:1353784, 2024; doi: 10.3389/fmed.2024.1353784

Background

Atopic dermatitis (AD) is a chronic inflammatory skin condition, affecting 15-30% of children and 2-10% of adults. Although AD is linked to sensitisation to various allergens, the role of the fungal microbiota (mycobiota) in disease severity remains poorly understood. The objective of this study was to examine the specific sensitisation towards the mycobiota in AD patients, with respective to their fungal skin colonisation and to analyse how the mycobiota impact the disease severity. The sensitisation patterns towards the two most common fungal microorganisms on human skin, Malassezia spp. and Candida albicans were investigated in 16 AD patients and 14 healthy controls (HC) over 18 years old in Switzerland using ALEX² and ImmunoCAP.

Key findings:

There was a significant difference in tIgE between AD and HC patients in both ALEX² and ImmunoCAP
There was a significant difference in M. sympodialis Mala s 6 sIgE (ALEX2) as well as C. albicans m 5 sIgE (ImmunoCAP) between HC and severe AD patients as well as between mild to moderate AD and severe AD
There was a statistically significant correlation between disease severity and sensitisation pattern: sensitisation of Malassezia spp. and C. albicans is increased in AD patients compared to HC
Malassezia spp. abundance decreased in severe AD compared to HC and mild to moderate AD
Candida spp. abundance increased in severe AD when compared to HC and mild to moderate AD

Conclusion:

This study revealed that AD, especially in severe cases, is linked to increased sensitisation towards the host’s mycobiota. Candida spp. skin colonisation positively correlates with AD. Malassezia spp. colonisation showed a negative correlation with AD, AD severity and sensitisation patterns.

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Tuten Dal S et al; Pediatr Allergy Immunol 35(2):e14093, 2024; DOI: 10.1111/pai.14093

Background:

Mites are common global aeroallergens. House dust mites (HDM) produce various molecular allergens (MA), which can elicit specific immune responses in sensitised individuals, causing distinct clinical symptoms. Understanding an individual's mite allergen sensitisation profile is essential for diagnosing and managing mite allergic diseases. The objective of this study was to investigate the MA sensitisation patterns among mite-sensitised children. For this purpose, 76 Turkish children with sensitisation to at least one mite MA were selected and the age-specific characteristics as well as potential cluster relationships among sensitisation patterns were investigated.

Key findings:

313 children were tested for their MA-sensitisation pattern by ALEX²
76 children showed sensitisation to at least one mite allergen molecule and were included in the study
69.7% of patients experienced co-sensitisation to other aeroallergens; namely 52.6% to tree pollen, 43.4% to grass pollen and 23.7% to weed pollen
Increasing co-sensitisation with other aeroallergens as patient’s age advanced
Most common sensitisations were to Der p 1/2 and Der f 1/2 (around 40%)
Der p 10/Blo t 10, Der p 20, Der p 23, and Gly d 2/Lep d 2 showed approximately 20% sensitisation rate
Der p 23 sensitisation rates and specific-IgE levels increased with age
Hierarchical clustering confirmed the strong correlations between Der f 2 and Der p 2, Der p 10 and Blo t 10, Der p 21 and Blo t 5, and Gly d 2 and Lep d 2
Der p 10 and Blo t 10-sensitive patients were mostly seen as mono sensitisation

Conclusion:

This study reveals an age-related increase in both the diversity and intensity of mite sensitisations in children, especially for Der p 23. Significant correlations among these sensitisations emphasises homologies among specific MAs. Furthermore, in the studied population a significant portion of mites sensitisation originated from tropomyosin and arginine kinase.

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Ukleja-Sokołowska N et al; Adv Dermatol Alergol 40(5):661-669, 2023; doi: 10.5114/ada.2023.132071

Background:

With shrimp sensitisation becoming more prevalent in developed countries, accurate diagnosis is critical. This requires selecting the most effective diagnostic method and tailoring the strategy to each patient’s unique clinical situation. This study aimed to investigate the allergen profile of shrimp-sensitised patients. For this purpose, 50 patients with positive prick-to-prick tests with tiger shrimp and with elevated specific IgE levels to shrimp extract in ImmunoCAP were studied using ALEX2, which allows to access specific IgE to shrimp allergen extracts and shrimp allergen components.

Key findings:

50 adult patients with positive prick-by-prick test and 35 patients with negative skin prick test (= control group) were included in the study
In the sensitised group, 22 patients were sensitised to at least one allergen component of Penaeus monodon, 20 to crab, 20 to lobster, 15 to Northern prawn and 12 to Shrimp mix (Litopenaeus setiferus, Farfantepenaeus aztecus, Farfantepenaeus dourarum)
The most prevalent shrimp allergen in the group of patients studied was tropomyosin, Pen m 1, followed by aginine kinase
Potential cross-sensitisations between shrimp tropomyosin Pen m 1 and Anisakis simplex tropomyosin Ani s 3 as well as Pen m 1 and American cockroach Per a 7 (Spearman’s Rank Correlation = 0.98)
41 patients were co-sensitised to at least one house dust mite allergen component
House dust mite (HDM) sensitisation was significantly higher in the shrimp sensitisation group
Most common HDM sensitisations were to Der p 2 (66%), Der p 23 (54%), Der p 5 (52%), and Der p 1 (52%)

Conclusion:

ALEX² offers a thorough overview of a patient's allergen profile because includes a largen number of shrimp allergen extracts and components. In this study, the most prevalent shrimp sensitisation was to shrimp tropomyosin (present in 34% of patients). This might imply that there are other shrimp allergen components responsible for sensitisation, which are not reflected in ALEX². Sensitisation to HDM is an important problem in shrimp-sensitised patients and patient sensitised to shrimp showed to have sensitisation also to other types of seafood.

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Walsemann T et al; Allergy 78(3):731-742, 2023; doi: 10.1111/all.15553

Background

House dust mites (HDM) are a major source of airborne allergens, affecting one in five EU citizens and significantly contributing to allergic asthma and atopic dermatitis. This study aimed to identify potential biomarkers by investigating the relationship between various HDM allergens and allergic rhinitis, asthma, and atopic dermatitis. The clinical relevance of nine HDM allergens (nDer p 1, rDer p 2, rDer p 5, rDer p 7, rDer p 10, rDer p 13, rDer p 20, rDer p 21, rDer p 23) were evaluated by analysing sensitisation patterns in 384 HDM-allergic adults exhibiting different clinical phenotypes.

Key findings:

Grouping of patients according to clinical phenotypes: single or combined presence of allergic asthma (AA), atopic dermatitis (AD) and allergic rhinitis (AR) ranked after sensitisation count and disease severity
The number of allergen sensitisation increased with clinical phenotype severity
Sensitisation to more than 3 HDM allergens associated with the occurrence of multiple allergic diseases
Sensitisation to more than 3 HDM allergens significantly associated with allergic asthma and/or atopic dermatitis
Sensitisation to 3 or less HDM allergens mainly associated with AR
AA patients were more often sensitised to Der p 5 and 21 compared to AR
AD patients were more often sensitised to Der p 2, 5, 10, 13, 20, and 21
Der p 20 – a biomarker for severe AD: Der p 20-IgE > 80 kU/L strongly associated with severe AD in 75% of patients

Conclusion:

This study underscores the clinical importance of both the total sensitisation count and specific allergens such as Der p 5, 20, and 21 which, are not available for routine diagnostics. Incorporation of those allergens in routine diagnostics along with the sensitisation count should enhance the diagnosis and risk assessment of HDM-allergic patients. In addition, the study concludes that HDM-multiplex test is a good diagnostic tool and carries an advantage over ImmuoCAP by requiring only a small amount of serum per analysis.

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