{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,4,29]],"date-time":"2026-04-29T23:09:52Z","timestamp":1777504192591,"version":"3.51.4"},"reference-count":55,"publisher":"MDPI AG","issue":"7","license":[{"start":{"date-parts":[[2025,3,26]],"date-time":"2025-03-26T00:00:00Z","timestamp":1742947200000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"name":"Italian Ministry of Health","award":["RF-2018-12367986"],"award-info":[{"award-number":["RF-2018-12367986"]}]},{"name":"Project RC IZS SI 02\/20","award":["RF-2018-12367986"],"award-info":[{"award-number":["RF-2018-12367986"]}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["IJMS"],"abstract":"<jats:p>The diagnosis of allergic reactions to Anisakis remains challenging due to the lack of specific allergens available for routine clinical use. However, the latest version of the multiplex macroarray ALEX-2 now allows the detection of specific IgE against Ani s 1, the major species-specific allergen, as well as Ani s 3 (tropomyosin), a highly cross-reactive molecule with homologs in other allergenic sources. This study aimed to evaluate the potential role of ALEX-2 in diagnosing Anisakis sensitization by comparing it with a previously validated diagnostic algorithm. Serum samples from patients with suspected Anisakis sensitization were consecutively collected at an Italian allergy centre. Diagnosis was based on a history of allergic reactions following seafood consumption, along with negative test results for fish allergy. All patients underwent skin prick testing and specific IgE measurement for Anisakis (p4), Ascaris (p1), shrimp (f24), and Dermatophagoides pteronyssinus (d1), as well as tropomyosins from house dust mites (d205) and shrimp (f351). Additionally, the basophil activation test (BAT) using crude Anisakis extract was performed. Patients were also tested using the ALEX-2 allergy macroarray. Correlation analyses and multiple logistic regression models were applied to assess associations between conventional singleplex tests and the severity of clinical manifestations. A total of 33 eligible subjects were recruited, including 20 females (60.6%). Seven (21.2%) were aged 0\u201329 years, eleven (33.3%) were 30\u201360 years old, and fifteen (45.5%) were over 60 years old. ALEX-2 showed positivity for Ani s 1 or Ani s 3 in 39.39% (95% CI: 22.90\u201357.86%) of subjects with confirmed Anisakis sensitization. A significant correlation was observed between Ani s 3 (r = 0.31 [95% CI: 0.04\u20130.56], p = 0.01) and Ascaris (r = 0.35 [95% CI: 0.129\u20130.55], p = 0.004) levels and the severity of clinical reactions. Despite the limitations of this cross-sectional study, including a small sample size, our preliminary findings suggest that the ALEX-2 macroarray may not be sufficiently sensitive for the first-line diagnosis of Anisakis allergy. However, it could provide valuable additional information, as Ani s 1 positivity indicates primary sensitization to the nematode, while Ani s 3 positivity appears to correlate with clinical severity. Larger prospective longitudinal studies are needed to confirm these findings and further assess the predictive value of ALEX-2 in diagnosing Anisakis allergy.<\/jats:p>","DOI":"10.3390\/ijms26073033","type":"journal-article","created":{"date-parts":[[2025,3,26]],"date-time":"2025-03-26T09:13:26Z","timestamp":1742980406000},"page":"3033","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":2,"title":["An Assessment of the Currently Available Molecular Assay for the Diagnosis of Anisakis Sensitization"],"prefix":"10.3390","volume":"26","author":[{"ORCID":"https:\/\/orcid.org\/0009-0005-9500-119X","authenticated-orcid":false,"given":"Maria","family":"Barrale","sequence":"first","affiliation":[{"name":"U.O.C di Patologia Clinica Ospedale Buccheri La Ferla FBF, 90123 Palermo, Italy"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-5090-1366","authenticated-orcid":false,"given":"Walter","family":"Mazzucco","sequence":"additional","affiliation":[{"name":"U.O.C. di Epidemiologia Clinica con Registro Tumori Azienda Ospedaliera Universitaria Policlinico \u201cPaolo Giaccone\u201d, 90127 Palermo, Italy"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Santo","family":"Fruscione","sequence":"additional","affiliation":[{"name":"U.O.C. di Epidemiologia Clinica con Registro Tumori Azienda Ospedaliera Universitaria Policlinico \u201cPaolo Giaccone\u201d, 90127 Palermo, Italy"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-5311-1359","authenticated-orcid":false,"given":"Maurizio","family":"Zarcone","sequence":"additional","affiliation":[{"name":"U.O.C. di Epidemiologia Clinica con Registro Tumori Azienda Ospedaliera Universitaria Policlinico \u201cPaolo Giaccone\u201d, 90127 Palermo, Italy"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Vincenzo","family":"Cantisano","sequence":"additional","affiliation":[{"name":"U.O.C di Patologia Clinica Ospedale Buccheri La Ferla FBF, 90123 Palermo, Italy"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-6300-6571","authenticated-orcid":false,"given":"Gaetano","family":"Cammilleri","sequence":"additional","affiliation":[{"name":"Istituto Zooprofilattico Sperimentale della Sicilia, 90129 Palermo, Italy"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Antonella","family":"Costa","sequence":"additional","affiliation":[{"name":"Istituto Zooprofilattico Sperimentale della Sicilia, 90129 Palermo, Italy"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Vincenzo","family":"Ferrantelli","sequence":"additional","affiliation":[{"name":"Istituto Zooprofilattico Sperimentale della Sicilia, 90129 Palermo, Italy"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Rosa","family":"Onida","sequence":"additional","affiliation":[{"name":"U.O.C di Patologia Clinica Ospedale Buccheri La Ferla FBF, 90123 Palermo, Italy"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-9391-9168","authenticated-orcid":false,"given":"Enrico","family":"Scala","sequence":"additional","affiliation":[{"name":"UOS di Allergologia Molecolare Clinica e di Laboratorio Istituto Dermatologico dell\u2019Immacolata, IDI-IRCCS, 00167 Rome, Italy"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Danilo","family":"Villalta","sequence":"additional","affiliation":[{"name":"SC Immunologia e Allergologia di Laboratorio Ospedale Santa Maria degli Angeli, 33170 Pordenone, Italy"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-4158-8333","authenticated-orcid":false,"given":"Carina Gabriela","family":"Uasuf","sequence":"additional","affiliation":[{"name":"Istituto di Farmacologia Traslazionale (IFT), Consiglio Nazionale delle Ricerche (CNR), 90146 Palermo, Italy"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-6243-7732","authenticated-orcid":false,"given":"Ignazio","family":"Brusca","sequence":"additional","affiliation":[{"name":"U.O.C di Patologia Clinica Ospedale Buccheri La Ferla FBF, 90123 Palermo, Italy"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"1968","published-online":{"date-parts":[[2025,3,26]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"93","DOI":"10.1016\/S0065-308X(08)60573-4","article-title":"Anisakis and anisakiasis","volume":"16","author":"Smith","year":"1987","journal-title":"Adv. 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