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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Barrale et al.; International Journal of Molecular Sciences, 2025; doi: 10.3390/ijms26073033

Background

Diagnosis of Anisakis allergy is challenging due to extensive cross-reactivity with shrimp, mites, and other nematodes. ALEX² is currently the only multiplex test offering molecular Anisakis allergens (Ani s 1 and Ani s 3). This study evaluated ALEX² as part of a diagnostic work-up, comparing it to a validated algorithm including ImmunoCAP and basophil activation test (BAT).

Key findings:

ALEX² detected Ani s 1 and/or Ani s 3 in 39.4% of patients with confirmed Anisakis sensitization, indicating limited sensitivity when used as a standalone first-line test.
Ani s 1 positivity on ALEX² indicates primary Anisakis sensitization, as Ani s 1 is a species-specific major allergen.
Ani s 3 (tropomyosin) positivity correlated with clinical reaction severity, aligning with tropomyosin’s known thermostability and involvement in severe reactions.
ALEX² helped differentiate primary sensitization vs. cross-reactive patterns, especially in patients with concomitant mite and crustacean sensitization.
Several patients positive for Anisakis extract were negative for both Ani s 1 and Ani s 3, highlighting the need for additional relevant molecules (e.g., Ani s 7, Ani s 13) to improve sensitivity.
ALEX² offered useful molecular context in complex cases, complementing BAT and ImmunoCAP, but should not replace first-line testing until more Anisakis allergens are included.

Conclusion:

ALEX² is not sensitive enough to serve as a stand-alone first-line test for Anisakis allergy. However, it provides clinically valuable molecular insights, particularly for confirming primary sensitization (Ani s 1) and assessing reaction severity patterns (Ani s 3). Expansion of the molecular panel is needed to enhance diagnostic accuracy.

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Bigas Peñuela et al.; J Investig Allergol Clin Immunol, 2026; doi: 10.18176/jiaci.1121

Background

Der p 23 is a major and clinically relevant house dust mite (HDM) allergen, yet many HDM extracts used for immunotherapy contain little or no standardized Der p 23. This study evaluated the prevalence of Der p 23 monosensitization and examined whether commercial extracts can inhibit Der p 23–specific IgE. ALEX² was used to exclude sensitizations to additional HDM molecules.

Key findings:

Among 1,700 HDM-allergic patients, 62.3% were sensitized to Der p 23, while 3.47% (59 patients) fulfilled strict ImmunoCAP criteria for Der p 23 monosensitization.
A subset of 22 patients was tested with ALEX², revealing that 27% were actually sensitized to additional mid-tier or minor HDM allergens (e.g., Der p 5, Der p 7, Der p 10, Der p 20), meaning true monosensitization was closer to 2.5%.
Clinical phenotype (rhinitis/asthma severity) did not differ between Der p 23–monosensitized and Der p 1/2-polysensitized groups.
ImmunoCAP inhibition assays showed wide variation between commercial HDM skin test extracts:
Extracts 1–4 inhibited Der p 23–IgE by ~70–80%,
Extract 5 (purified Der p 23) showed lower and more variable inhibition, likely due to structural or concentration-related factors.
Findings highlight that many extract-based products contain insufficient or variable Der p 23 content, raising concerns for AIT efficacy in this patient subgroup.

Conclusion:

True monosensitization to Der p 23 is uncommon but clinically relevant. ALEX² proved essential in ruling out hidden sensitizations, showing that one-quarter of presumed Der p 23–monosensitized patients were actually sensitized to additional HDM molecules. Commercial HDM extracts differ markedly in their ability to inhibit Der p 23–specific IgE, underscoring the need for improved standardization of Der p 23 in diagnostic and therapeutic formulations.

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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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Bourgoin-Heck et al.; Pediatric Allergy & Immunology, 2024; doi: 10.1111/pai.70014

Background

This study used ALEX² to characterize molecular sensitization profiles in 367 children with moderate to severe asthma living in the megacity region of Paris. The aim was to identify sensitization-driven asthma phenotypes using component-resolved cluster analysis.

Key findings:

ALEX² enabled detailed molecular clustering by analysing up to 300 allergens (extracts + molecules), revealing distinct sensitization profiles at preschool and school age.
In preschool children, three clusters emerged:
Few sensitizations with non-T2 inflammation;
Predominant house dust mite (HDM) sensitization;
Severe asthma with multiple inhalant + food sensitizations and high eosinophils.
In school-age children, five clusters were identified, including the classic HDM and PR-10 phenotypes, but also two severe forms: (4) Tropomyosin-driven severe asthma (novel finding), (5) Multiple sensitizations with very high total IgE and highest hospitalization rates.
Tropomyosin sensitization emerged as a new marker of severe asthma, associated with cockroach, storage mite, and HDM exposure patterns identifiable only through broad molecular arrays like ALEX².
PR-10–driven pollen–food sensitization was unexpectedly common in severe asthma at school age, likely influenced by climate and urban environmental factors.
ALEX²’s wide allergen coverage enabled detection of previously overlooked profiles, especially storage-mite and indoor-environment sensitizations that contribute to severe asthma.

Conclusion:

ALEX² allowed high-resolution mapping of sensitization profiles, revealing novel phenotypes, particularly tropomyosin-associated severe asthma, and highlighting the importance of molecular diagnostics in personalizing asthma assessment in children living in complex urban environments.

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Caka et al.; Pediatric Allergy & Immunology, 2025; doi: 10.1111/pai.70233

Background

Storage proteins (2S albumins, 7S vicilins, 11S legumins) are major drivers of persistent and severe food allergy. This study used ALEX² multiplex testing in 350 children to map molecular sensitisation patterns, evaluate cross-sensitisation, and identify markers predicting clinical reactivity.

Key findings:

ALEX² identified one large storage-protein cluster spanning peanut, tree nuts, sesame, poppy seed and buckwheat, driven mainly by cross-sensitisation, with contributions from co-sensitisation and molecular spreading.
Strongest correlations were seen between known clinical marker pairs (e.g., Ana o 3–Pis v 1, Jug r 4–Cor a 9, Ara h 2–Ara h 6), confirming established cross-reactivity patterns.
2S albumins were the most predictive molecules for clinical reactivity across allergens: Ara h 2 (peanut), Cor a 14 (hazelnut), Jug r 1 (walnut), Ana o 3 (cashew), Pis v 1 (pistachio), Ses i 1 (sesame) and Pap s 2S albumin (poppy seed).
Lower ALEX² sensitisation threshold (≥0.1 kUA/L) and exclusion of patients with very low total IgE improved diagnostic accuracy (higher AUC values).
Molecules with broad cross-sensitisation patterns (e.g., Cor a 9, Cor a 11, Jug r 4, Ana o 2) showed limited specificity and should be interpreted with caution.
α-hairpinins Mac i 2S and Pap s 2S albumin showed fewer correlations and may represent more specific markers; Pap s 2S albumin appears to be a novel poppy seed allergy marker.
Authors note that use of purified natural allergens may introduce trace cross-contamination (important for Ara h 1/3 and Pis v 2/3), recommending recombinant molecules in future platforms.

Conclusion:

ALEX² enables high-resolution mapping of storage-protein sensitisation, supporting differentiation between cross- and co-sensitisation and improving prediction of clinical reactivity. 2S albumins remain the most informative markers for food allergy risk. Interpretation benefits from considering total IgE and low-level sensitisation, making ALEX² a valuable tool in complex paediatric food allergy diagnostics.

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Čelakovská et al.; Journal of Immunotoxicology, 2025; doi: 10.1080/1547691X.2025.2561586

Background

CD200 is an immunoregulatory molecule that transmits inhibitory signals and helps control inflammation. Its relationship to IgE-mediated allergy is poorly understood. This study examined whether CD200 expression on B-lymphocyte subsets correlates with specific IgE to molecular allergens, measured using ALEX², in atopic dermatitis (AD) patients with and without dupilumab therapy.

Key findings:

ALEX² detected sensitization to storage mite (Gly d 2, Lep d 2), pet (Fel d 1, Can f 1/2), mold (Asp f 6, Mala s 11, Alt a 1/6, Mala s 6, Cla h), shrimp (Pen m 2), and cockroach (Bla g 9) components in both AD groups.
In patients without dupilumab, correlations between CD200 expression and specific IgE were very low (typically R² < 10%), with the highest association seen for Fel d 1 (10.1%).
In dupilumab-treated patients, correlation values were frequently negative, suggesting an inverse or non-linear relationship between CD200 expression and IgE levels.
Dupilumab-treated patients showed higher mean IgE levels for some components (e.g., Bla g 9, Mala s 11), though variability prevented consistent statistical significance.
Results contrast with previous CD23 studies from the same group, where dupilumab produced strong CD23–IgE associations; here, CD200 did not behave similarly, indicating distinct immunoregulatory pathways.
Overall, no meaningful association between CD200 expression and molecular IgE patterns was confirmed.

Conclusion:

Using ALEX² molecular IgE data, the study demonstrates that CD200 expression on B-lymphocytes does not correlate with sensitization profiles in AD patients, regardless of dupilumab treatment. CD200 therefore appears unlikely to play a central role in IgE regulation for these allergen families, helping narrow the search toward other mechanisms such as CD23- or FcεRI-related pathways.

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Čelakovská et al.; Journal of Immunotoxicology, 2025; doi: 10.1080/1547691X.2025.2507311

Background

CD23 (FcεRII) is a low-affinity IgE receptor involved in IgE regulation, but its relationship to molecular sensitization patterns in atopic dermatitis (AD) is unclear. This study measured specific IgE using ALEX² Allergy Xplorer and evaluated how CD23 expression correlates with IgE to individual pollen molecules in AD patients with and without dupilumab therapy.

Key findings:

ALEX² identified high rates of sensitization to Bet v 1, Cor a 1.0103/1.0401, Phl p 1, Lol p 1, and Cyn d across both groups, with overall higher IgE levels in dupilumab-treated patients.
Dupilumab-treated patients showed the strongest associations between CD23 expression and IgE to Bet v 1, Cor a 1.0103, Cor a 1.0401, and Phl p 1 (R² values ~21–35% across B-cell subsets).
Without dupilumab, the strongest associations occurred with Cyn d (Bermuda grass), likely reflecting cross-reactivity with other β-expansin grasses present in Central Europe.
The pattern of CD23–IgE interaction differed by molecular component, indicating allergen-specific variability in how IgE engages CD23.
For Phl p 5, associations were weak or non-linear, likely due to its strong basophil-activating capacity and high IgE-binding epitope density, reducing the role of CD23-mediated uptake.
Findings suggest that dupilumab may shift IgE biology toward greater IgE–CD23 binding and reduced FcεRI involvement, a mechanism that could contribute to its anti-inflammatory effects.

Conclusion:

Using ALEX², the study demonstrates that dupilumab alters how IgE to specific pollen molecules interacts with CD23 on B-lymphocytes. Stronger CD23–IgE associations for Bet v 1, Cor a 1.0103/1.0401, and Phl p 1 under dupilumab may reflect a shift toward non-inflammatory IgE handling. These insights may help explain variable dupilumab responses and guide more personalized evaluation of molecular sensitization in AD.

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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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Didziokaite et al; Frontiers in Medicine, 2024; doi: 10.3389/fmed.2024.1403477

Background

Human seminal plasma (HSP) allergy is an extremely rare type I hypersensitivity reaction that may contribute to unexplained infertility. Fewer than 100 cases have been documented globally, and awareness among clinicians remains low. This case report describes a 29-year-old woman with unexplained infertility, respiratory allergies, and eosinophilia who was ultimately diagnosed with HSP allergy following positive skin-prick testing with her partner’s seminal plasma.

Key findings:

The patient exhibited long-standing allergic disease (asthma, rhinitis) and eosinophilia, along with local and systemic reactions after unprotected intercourse.
ALEX² testing revealed strong sensitization to multiple inhalant allergens, including Can f 5, a prostatic kallikrein from male dogs known to cross-react with human PSA proteins.
The identification of marked Can f 5 sensitization prompted targeted investigation for HSP allergy.
Skin prick testing with her partner’s seminal plasma produced a positive wheal equal to histamine, confirming IgE-mediated seminal plasma allergy.
Follow-up testing showed molecular spreading over time and further increase in Can f 5 levels, correlating with worsening symptoms after semen exposure.
Treatment options discussed included barrier contraception, pre-intercourse antihistamines, and sperm desensitization, though the latter was not available locally.
The patient had concomitant asthma and mild endometriosis, both of which can contribute to systemic inflammation and may negatively influence fertility outcomes.

Conclusion:

This case highlights the diagnostic value of molecular allergy testing, particularly Can f 5, in revealing otherwise overlooked seminal plasma allergy in women with unexplained infertility. Increased clinical awareness is needed, as timely identification may guide appropriate counselling, targeted management, and reproductive planning.

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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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González-Pérez et al.; Nutrients, 2025; doi: 10.3390/nu17091405

Background

Edible insects (EIs) are emerging as novel food sources, but their allergenic risks remain unclear, particularly in subtropical regions with high exposure to house dust and storage mites. This study used ALEX² to characterize IgE sensitization patterns to edible insects and related pan-allergens, evaluating cross-reactivity with mites and crustaceans.

Key findings:

Among 634 patients tested by ALEX², 21.76% were sensitized to at least one edible insect extract (house cricket, locust, or mealworm).
Tropomyosin was the dominant pan-allergen (63.76%), followed by troponin-C, arginine kinase, and other muscle proteins, confirming strong cross-reactivity with mites and crustaceans.
95.66% of edible-insect–sensitized individuals were also sensitized to at least one mite molecule; group 2 mite allergens (Der p 2, Der f 2) and Der p 23 were most common.
Notably, 23.18% of EI-sensitized subjects showed no pan-allergen reactivity on ALEX², suggesting additional insect-specific allergens or alternative sensitization pathways.
In patients with only respiratory allergy (no food allergy), EI sensitization was entirely linked to robust mite sensitization, while pan-allergen reactivity was rare.
Strong correlations were observed between EI sensitization and TM/AK homologues (e.g., Der p 10, Blo t 10), supporting arthropod cross-reactivity as a major driver.

Conclusion:

ALEX²-based profiling shows that edible insect sensitization in subtropical climates is strongly shaped by regional mite exposure and cross-reactive muscle proteins such as tropomyosin. A subset of patients appears to develop sensitization beyond known pan-allergens, highlighting the need for further molecular characterization. EI sensitization should be considered a relevant emerging allergenic phenomenon rather than incidental IgE binding.

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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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Heidari et al.; Frontiers in Allergy, 2025; doi: 10.3389/falgy.2025.1674423

Background

Crustacean allergy is a major cause of food-induced anaphylaxis in tropical regions. Strong epidemiological links exist between crustacean and house dust mite (HDM) allergy, but the molecular basis of this relationship is incompletely understood. This study analyzed ALEX2 molecular IgE profiles in well-characterized crustacean-allergic individuals to identify cross-reactive and co-sensitization patterns.

Key findings:

ALEX² detected mite sensitization in 85% of crustacean-allergic individuals, highlighting substantial overlap between the two allergy groups.
The most frequent mite components were Lep d 2 (64.8%), Der p 23 (61.1%), Der f 2 (59.2%), and Der p/f 1 (55–57%) (Figure 1, p.3), showing that major HDM allergens dominate co-sensitization.
Tropomyosin (Pen m 1) sensitization strongly correlated with IgE to mite tropomyosins Der p 10 and Blo t 10 (p < 0.0001), confirming classical cross-reactivity.
Non-tropomyosin molecules also played a major role: Pen m 2 (arginine kinase) sensitization correlated with Der p 20 IgE (p < 0.0001), while Cra c 6 (troponin C) was associated with Der p 5, Der p 20, Der p 23, Gly d 2, and Lep d 2.
ALEX² crustacean-negative subjects still showed strong mite sensitization (Der p 1: 50%; Der p 2: 42.9%), suggesting primary mite sensitization with secondary clinical reactions to crustaceans.
Among patients with anaphylaxis, Der p 10, Der p 23, and Lep d 2 sensitization were most strongly associated with severe systemic symptoms, indicating potential risk markers.

Conclusion:

ALEX² profiling reveals that in tropical regions, crustacean allergy is shaped not only by tropomyosin cross-reactivity but also by non-tropomyosin allergens, especially arginine kinase, troponin C, group 2 mite allergens, and Der p 23. These findings underline the clinical importance of component-resolved diagnostics in distinguishing cross-reactivity from co-sensitization and in assessing risk for severe reactions.

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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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Lupinek et al; Int. J. Mol. Sci., 2025; doi: 10.3390/ijms26094249

Background

ALEX² is a globally used multiplex IgE macroarray. With nearly 400,000 real-world test results available by 2023, MADx applied a data-driven approach to redesign the allergen panel for ALEX³. The goal was to improve diagnostic sensitivity, increase global exposome coverage, and refine the balance between extracts and defined allergen molecules.

Key findings:

For spreading pellitory, many sera were exclusively extract-positive and not explained by pan-allergens or CCDs, so the Parietaria extract was retained alongside Par j 2 to capture additional clinically relevant epitopes.
For ribwort plantain, Pla l 1 and the extract showed strong co-reactivity with very few true extract-only positives, so the Pla l extract was removed while Pla l 1 was kept as a specific marker.
Storage protein families (2S, 7S, 11S) showed frequent monosensitisation and diverse co-reactivity patterns, preventing the definition of surrogate markers; therefore all storage proteins were kept and further expanded on ALEX³.
In contrast, PR-10 allergens were highly redundant and largely covered by Bet v 1; several PR-10s were removed, while regionally important markers like Aln g 1 and Que a 1 were retained or added.
For Ole e 1-like proteins, limited co-reactivity and many monosensitisations indicated source-specific epitopes; thus all four existing Ole e 1-like allergens were kept and Sal k 5 was added.
Profilin analysis showed that Phl p 12 and Cuc m 2 sufficiently represent the profilin family, allowing removal of four redundant profilins without losing diagnostic coverage.
For grasses, a refined combination of extracts and components (Phl p 1, Cyn d 1, Pas n extract, Zea m 1) defined, while redundant markers (e.g. Lol p 1, Bermuda extract) were removed, optimising sensitivity and reducing overlap.
52 new allergen molecules and 1 extract were added to ALEX³ including clinically important markers such as alpha-gal, seafood allergens (Lit v 7, Mac r 1/2), chicken meat allergen Gal d 7, banana/mango allergens (Mus a 2, Mus a 5, Man i 1), wheat markers (Tri a 36, Tri a 37) new pan-allergens (Bet v 7, Mala s 13), and venom markers (Api m 2, Dol m 2, Dol m 5).

Conclusion:

Using big data from nearly 400,000 ALEX² tests enabled a systematic refinement of the multiplex IgE array. ALEX³ enhances diagnostic accuracy by expanding clinically relevant molecular markers, reducing redundancy, and improving coverage of region-specific exposomes.

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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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Murray et al; S Afr J Child Health, 2024; doi: 10.7196/SAJCH.2024.v18i4.2234

Background

Molecular allergy diagnostics allow clinicians to distinguish true primary sensitisation from cross-reactivity. This study compared performance of ALEX² with the ISAC assay across food, inhalant and cross-reactive allergen components using 100 patient sera.

Key findings:

ALEX² and ISAC showed strong overall agreement: 58% of components had very good Kappa values, 33% good, and only 8% moderate.
Highest agreement was observed for Der p 23, Der f 2, Tri a 19 and Asp f 1 (Kappa 0.98–1.00), demonstrating excellent concordance for key clinically relevant allergens.
Components with moderate agreement were primarily food allergens (Gly m 4, Ara h 8, Gly m 5, Tri a A/TI), reflecting known variability in PR-10 and certain storage proteins.
ALEX² showed high analytical specificity due to CCD inhibition, reducing false positives compared to ISAC (as also illustrated by Fig. 2 on page 178).
Positive percent agreement exceeded 80% for ~90% of all components, and negative percent agreement was >90% for nearly all allergens tested.
For inhalant allergens, ALEX² performed extremely well, with very good agreement for 73% of components, including mites, pets, grasses, moulds and pollen.
Cross-reactive markers (profilin, LTP, Bet v 1 family) showed good–very good concordance, though PR-10 proteins displayed expected variability across platforms.
Clinical questionnaire data linked sensitisation patterns detected by ALEX² to symptoms such as atopic dermatitis, food reactions and anaphylaxis, reinforcing the clinical relevance of molecular diagnostics.

Conclusion:

The study demonstrates a high degree of concordance between ALEX² and ISAC for detecting IgE to allergen components. ALEX² offers broader molecular coverage, the added benefit of CCD inhibition, and simultaneous measurement of extracts and components, providing a comprehensive and clinically informative sensitisation profile.

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Nösslinger et al.; JDDG – Journal der Deutschen Dermatologischen Gesellschaft, 2025; doi: 10.1111/ddg.15810

Background

This study investigated how sensitization to stable (2S albumins, 7/8S & 11S globulins) versus unstable (PR-10, profilins) allergen families correlates with epithelial barrier dysfunction and clinical symptoms. ALEX² was used to define detailed molecular sensitization patterns in 216 patients from South Tyrol.

Key findings:

ALEX² identified distinct sensitization groups, enabling comparison of stable vs. unstable allergen profiles across age groups and disease manifestations.
Patients sensitized to stable food allergens were significantly younger and showed more eczema, higher IGA eczema scores, and increased TEWL values, markers of epithelial barrier impairment.
Stable-allergen sensitization was strongly associated with combined gastrointestinal + cutaneous symptoms, whereas unstable-allergen sensitization more often presented with isolated respiratory symptoms.
Patients co-sensitized to both stable and unstable allergens had the highest FeNO values and increased rates of bronchial asthma, indicating greater airway inflammation.
Results support the concept that early epithelial barrier dysfunction promotes sensitization to stable allergens, which in turn increases risk for later asthma when aeroallergen co-sensitizations develop.
Authors highlight that multiplex assays such as ALEX² are particularly suitable for identifying complete sensitization patterns in patients at risk of multi-system involvement and asthma development.

Conclusion:

ALEX² effectively differentiates sensitization to stable vs. unstable allergen families, providing clinically relevant insights into epithelial barrier dysfunction and asthma risk. Multiplex profiling can support early identification of high-risk patients and guide tailored management strategies.

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Obermeyer et al.; Contact Dermatitis, 2025; doi: 10.1111/cod.70008

Background

Compositae (Asteraceae) plants frequently cause allergic contact dermatitis (ACD) in florists due to sesquiterpene lactones. Immediate-type hypersensitivity to lettuce is rare. This case report describes a florist with chronic occupational ACD and episodes of anaphylaxis after eating lettuce and radicchio. Specific IgE was evaluated using ImmunoCAP and multiplex arrays including ALEX.

Key findings:

The patient showed strong positive patch tests to Compositae mix, sesquiterpene lactone mix, and parthenolide, confirming occupational ACD.
Rub tests with fresh lettuce (butterhead and romaine) triggered immediate wheal formation, supporting IgE-mediated reactivity despite negative commercial IgE tests.
ImmunoCAP and ALEX multiplex testing detected no IgE sensitization to known plant and food allergens, including LTPs, profilins, PR-10 proteins, and lettuce extract (f215).
A separate FOX multiplex array detected low-level IgE to chicory, a related Compositae species, suggesting possible cross-reactivity.
Known lettuce allergens include Lac s 1 (nsLTP), thaumatin-like protein, aspartyl protease, profilin, and recently described EP1-like and kirola-like proteins—but none were detected in this patient.
Authors propose sensitization to an unknown, not yet characterized Compositae protein, facilitated by chronic occupational skin barrier damage.

Conclusion:

Despite negative conventional IgE tests, including ALEX, the clinical picture supported immediate-type lettuce allergy. Chronic occupational exposure to Compositae plants likely induced both delayed-type ACD and immediate sensitization to an unidentified lettuce-related protein. This case highlights the limits of current molecular panels for rare plant allergens and the importance of fresh material testing in diagnostic evaluation.

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Özdemiral et al.; Pediatric Allergy and Immunology, 2025; doi: 10.1111/pai.70157

Background

Serum albumins (SAs) are heat-labile, highly cross-reactive animal proteins found in milk, meat, dander and epithelial secretions. This study examined SA sensitisation patterns—including Bos d 6—and their relevance to red meat reactions in children with cow’s milk allergy (CMA), using ALEX² molecular profiling.

Key findings:

61.4% of children with CMA were sensitised to Bos d 6, and ~40% of these were Bos d 6 monosensitised.
ALEX² identified frequent co-sensitisation to other mammalian SAs: Sus s 1 (47%), Equ c 3 (26%), Fel d 2 (23%), and Can f 3 (16%). Bos d 6 showed strong correlation with Sus s 1 (r = 0.79) and moderate correlation with Equ c 3 and Can f 3 (heatmap on page 5).
Despite high sensitisation rates, clinically relevant red meat reactions were uncommon (10%), mostly triggered by raw or undercooked meat, contact with raw meat/blood, or inhalation of steam.
All red-meat-reactive patients were Bos d 6 positive, supporting its role as the key molecular risk marker.
Children universally tolerated well-cooked red meat, consistent with heat-induced loss of SA allergenicity.
Sensitisation to other milk allergens (Bos d 4, Bos d 5, Bos d 8) decreased after CMA resolution, but Bos d 6 persisted in 67% of tolerant patients, indicating slower decline.
Hierarchical clustering (page 6) showed Bos d 6 grouping closely with Sus s 1, Equ c 3 and Can f 3, while Gal d 5 clustered separately, confirming limited cross-reactivity with poultry albumin.
Avoidance behaviour was common: 18% had long-term beef elimination diets, despite tolerating cooked meat, suggesting over-avoidance rather than true clinical risk.

Conclusion:

ALEX² revealed that Bos d 6 sensitisation is common in children with CMA but rarely translates into clinically significant red-meat allergy, particularly when meat is well cooked. Molecular profiling with ALEX² enables identification of cross-sensitisation patterns, supports risk stratification, and helps prevent unnecessary dietary avoidance in paediatric CMA patients.

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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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Rivas-Juesas et al; Allergol Immunopathol (Madr), 2025; doi: 10.15586/aei.v53i3.1298

Background

Environmental exposures in Mediterranean climates strongly influence paediatric allergy development. This study characterised molecular sensitisation patterns in children with allergic rhinitis (with or without asthma) using ALEX² to identify clinically relevant molecules and support precision immunotherapy.

Key findings:

ALEX² revealed high sensitisation to house dust mites (74.5%), with Der p 23 being the most frequently recognised component (59.6%), followed by group 2 (Der p 2 / Der f 2) and group 1 allergens.
Polysensitisation was common (66%), with the most frequent overlap involving mites, pollen, and animal epithelia.
Pollen sensitisation (57.4%) was dominated by Ole e 1 and Ole e 9 for olive, and Phl p 1 for grasses, consistent markers of genuine sensitisation in Mediterranean regions.
Sensitisation to animal epithelia was detected in 44.6%, with Fel d 1 and Can f 1 as the leading molecular allergens; older children (11–15 years) showed the highest IgE responses.
Fungal sensitisation was less common, though Alt a 1 was notable in children <5 years, indicating early onset in this region.
The 6–10 year age group showed the highest overall IgE levels, suggesting a peak window of sensitisation intensity.
The study highlights that several diagnostic extracts lack key molecules (e.g., Der p 23, Ole e 1), reinforcing that component-resolved diagnostics via ALEX² increase diagnostic accuracy and guide more precise allergen immunotherapy formulations.

Conclusion:

ALEX² molecular profiling demonstrated that mite, olive, grass and cat allergens are the dominant sensitisation drivers in Mediterranean children, with early onset and frequent polysensitisation. Incorporating clinically relevant molecular components into diagnostic and therapeutic extracts is essential for effective personalised allergy management.

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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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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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Scala et al.; Allergy, 2024; doi: 10.1111/all.16377

Background

Atopic dermatitis (AD) may involve IgE autoreactivity against human-homologous proteins. Using ALEX2, the authors examined IgE responses to 183 allergen molecules to identify human-homologous exogenous molecular allergens (HEMAs) and developed an IgE-molecular mimicry index (IgE-MMI). They assessed whether IgE to HEMAs or the IgE-MMI correlate with AD severity, clinical phenotypes, and response to dupilumab.

Key findings:

48 ALEX2 allergen molecules showed significant sequence similarity to human proteins, defining them as HEMAs. These included arginine kinase, enolase, lipocalins, cyclophilins, and MnSOD.
IgE to HEMAs was strongly associated with severe AD, far more than IgE to non-HEMA molecules (only 3/135 non-HEMAs showed any association).
Key HEMAs linked to severe AD were arginine kinase (Bla g 9, Der p 20, Pen m 2), enolase (Asp f 6), MnSOD (Alt a 6, Mala s 11), lipocalins (Can f 1, Fel d 7), and cyclophilin (Mala s 6).
The IgE-MMI was positive in 53.7% of severe AD patients, but only 10.7% of non-AD allergic controls and 20–40% of milder AD forms.
Distinct AD phenotypes showed different HEMA-IgE patterns: flexural and generalized eczema correlated most strongly with MnSOD and lipocalin reactivity; head-and-neck dermatitis correlated with MnSOD.
Dupilumab non-responders showed strong IgE to enolase, while responders more often had IgE to MnSOD and NPC-2 (Der p 2 / Gly d 2 family), suggesting predictive biomarker potential.

Conclusion:

ALEX²-based molecular profiling reveals that IgE to HEMAs, proteins with structural similarity to human molecules, is highly enriched in severe AD. The newly proposed IgE-MMI may help identify AD severity, distinguish clinical phenotypes, and predict dupilumab response. Microarray-based component testing appears essential for detecting these complex autoreactivity-linked IgE patterns.

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Scala et al.; Allergy, 2025; doi: 10.1111/all.70114

Background

Thaumatin-like proteins (TLPs, PR-5 family) are structurally stable plant proteins found in fruits such as kiwi (Act d 2) and apple (Mal d 2). Their clinical relevance is unclear, and they are often overshadowed by stronger panallergens. This study used the ALEX² multiplex platform to analyze prevalence, co-sensitization patterns and symptom relevance in a large Italian cohort (n=7176).

Key findings:

TLP sensitization was rare (1.9%), predominantly to Act d 2; Mal d 2 was much less frequent, and cross-recognition between the two was minimal (<6%). Isolated TLP sensitization was very uncommon (14.6%) and mostly clinically silent,16/20 mono-sensitized individuals had no symptoms, and others had only mild reactions.
85% of TLP-positive patients were co-sensitized to major plant panallergens (nsLTP, PR-10, profilin, seed storage proteins), indicating that TLP IgE appears mostly within broader sensitization profiles.
Clinical symptoms in co-sensitized patients (oral allergy syndrome, urticaria, anaphylaxis, FDEIA) were largely attributable to coexisting panallergens, not TLPs themselves.
Quantitative TLP-specific IgE showed poor predictive value for clinical severity (Act d 2 AUC 0.51–0.56), even after adjusting for age, sex, and co-sensitizations. Findings confirm that TLP-specific IgE alone is not a reliable diagnostic or prognostic marker, emphasizing the need for integrated molecular interpretation.

Conclusion:

Using ALEX², the study demonstrates that TLP sensitization is infrequent and usually clinically insignificant when isolated. Relevant symptoms arise mainly from co-sensitization to stronger panallergens, not from TLPs themselves. TLP IgE levels have limited diagnostic value, reinforcing the importance of multiplex molecular profiling to correctly contextualize emerging plant allergens.

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Scala et al.; Allergy, 2025; doi: 10.1111/all.70062

Background

Wheat allergy comprises distinct sensitization pathways to water-soluble proteins (e.g., nsLTP Tri a 14, α-amylase inhibitor Tri a 30) and gluten-derived allergens (e.g., ω-5-gliadin Tri a 19). Cross-reactivity with other cereals is poorly defined. This study analyzed sequence homology and IgE profiles across 17,510 patients, including a clinically evaluated cohort of 7,201 individuals tested with ALEX².

Key findings:

Tri a 14 (nsLTP) was the predominant wheat allergen, representing 64% of wheat sensitizations; Tri a 19 and Tri a 30 were less frequent (18% and 23%).
ALEX² data showed distinct and largely non-overlapping sensitization pathways: soluble allergens (Tri a 14, Tri a 30) and gluten-derived allergens (Tri a 19) rarely co-occurred.
Tri a 14 and Tri a 19 independently correlated with severe reactions, including wheat-dependent exercise-induced anaphylaxis (WDEIA), highlighting their diagnostic importance.
Sequence analyses revealed broad cross-reactivity potential for Tri a 14, including unexpected high similarity (72%) with a previously uncharacterized maize nsLTP (Zea m B6SIF2), whereas Tri a 19 was restricted to gluten-containing cereals (wheat, rye, barley, spelt).
Clinical ALEX² data showed different risk profiles:
Gluten-containing cereals → increased WDEIA risk
Gluten-free cereals (rice, maize, millet) → more severe systemic reactions
Pseudocereals (quinoa, amaranth, buckwheat) → typically mild to moderate symptoms
Sensitization to Tri a 30 and Tri a 14 was also associated with asthma, suggesting links to respiratory comorbidity.

Conclusion:

ALEX² revealed that wheat allergy consists of two divergent molecular sensitization pathways—one targeting soluble proteins and another directed toward gluten-derived molecules. Tri a 14 and Tri a 19 emerged as major markers for severe reactions, including WDEIA. Cross-reactivity across cereals varies significantly by allergen family, underscoring the need for expanded molecular panels and component-resolved diagnostics to improve dietary guidance and risk stratification.

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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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SIAIP Allergy Diagnostic Commission; Italian Journal of Pediatric Allergy and Immunology, 2024; doi: 10.53151/2531-3916/2024-436

Background

This SIAIP commission document reviews the technical principles, analytical performance, and clinical use of multiplex IgE platforms, ISAC and ALEX² emphasizing strengths, limitations, and appropriate clinical indications for molecular diagnostics.

Key findings:

ALEX² offers the broadest analyte coverage, combining 178 molecules + 117 extracts + total IgE, whereas ISAC measures 112 molecules and no extracts.
Extract-based IgE performance of ALEX² correlated moderately with ImmunoCAP (κ = 0.64 for inhalants, κ = 0.47 for foods), reflecting the expected semi-quantitative nature of all multiplex assays.
For molecular components, studies comparing ALEX² vs. ISAC report good qualitative agreement, but quantitative correlations vary due to different technologies, measurement ranges, and calibration systems.
ALEX² uses CCD inhibition via the sample diluent, reducing false positives; however, inhibition efficiency ranged between 60–95% depending on the study.
A known limitation of ALEX² is a reduced upper measurement range, with signal plateauing above ~35–50 kUA/L, affecting quantitative correlation with ISAC and singleplex assays.
Multiplex systems are valuable for complex polysensitisation, idiopathic anaphylaxis and difficult diagnostic cases, but can be harmful if used without specialist interpretation, risking overdiagnosis or unnecessary diet restrictions.
Both platforms are semi-quantitative and less analytically sensitive than singleplex assays, meaning numerical IgE values are not interchangeable across systems.

Conclusion:

ALEX² and ISAC provide powerful molecular profiles that support precision diagnosis when used appropriately. ALEX² stands out for its broader allergen panel, inclusion of extracts, and CCD inhibition, whereas both systems require specialist interpretation due to semi-quantitative output and methodological differences.

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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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